John M Woodley

Gürkan Sin

Krist V Gernaey

Introducing mechanistic models in Process Analytical Technology education ((Research Highlight))

Keywords: control, education, modeling, process analytical technology (PAT)

The Process Analytical Technology (PAT) guidance of the Food and Drug Administration (FDA) is designed to move pharmaceutical production away from off-line product quality testing to 'real-time release' strategies supported by online measurement of critical process variables. A successful PAT implementation primarily requires in-depth process knowledge but also a number of tools and methods including on-line analysis, process monitoring, as well as process control. In order to achieve successful PAT implementation, the students and professionals already working in the pharmaceutical industry will need more skills particularly within the design, tuning and implementation of process control algorithms. Based on reported experiences in other engineering disciplines such as chemical and environmental engineering, we conclude that development of a benchmark (mechanistic) modeling platform for pharmaceutical manufacturing systems is needed. Supported by a team of experts from academia and industry, this benchmarking effort can deliver documented and validated models of selected case-studies from pharmaceutical industry. Such publicly available and expert validated models could significantly contribute to educating students and professionals with skills that are essential for implementing the Process Analytical Technology guidance in pharmaceutical manufacturing systems.

Most pharmaceutical products today are small-molecule targets that are made using organic synthesis. In recent years the complexity of the molecules has increased in an attempt to improve selectivity and reduce side-effects. Higher molecular-weight molecules with multiple-chiral centers has also seen many processes now start to incorporate enzyme-based catalysis (driven also by environmental needs) [START_REF] Pollard | Biocatalysis for pharmaceutical intermediates: the future is now[END_REF]. Alongside these targets are also a growing group of biotechnology-based macromolecular targets, often referred to as biopharmaceuticals, such as insulin for example [START_REF] Johnson | Human insulin from recombinant DNA technology[END_REF][START_REF] Barfoed | Insulin production technology[END_REF]. Whether a small-molecule or a bio-pharmaceutical target is required, until a couple of years ago, all such manufacturing processes were fixed according to the specific production process description that was provided in the Drug Master File submitted to the FDA (or any other national regulatory body equivalent to the FDA).

It is common knowledge that there can exist considerable variation in raw materials, e.g. when switching from one batch of a chemical to the next batch, or when acquiring a chemical from an alternative supplier. Excessive variation in product quality -or in other words sub-optimal performance of the processcould be the result of having a fixed production process when confronted with unknown disturbances or variations in raw materials composition and/or properties, as schematically illustrated in the upper part of Figure 1. This does not affect the consumer (patient) directly, since time-consuming off-line laboratory testing of product quality is performed to ensure that every product batch achieves the required quality standards. The latter approach -also called the 'lab-centric' approach -is characterized by minimal closed loop, real-time control and limited enterprise-wide data availability. However, since each product batch of inferior quality will lead to increased production costs of a pharmaceutical, the price of the final product becomes higher as a consequence of these inefficient production methods. An additional factor that might lead to inefficient production processes is that there is usually very little time for production process development. Indeed, any delay in establishing a production process leads to decreased profit in view of the limited patent lifetime for each pharmaceutical, and thus process development by necessity is a compromise to ensure rapid time-to-market. During the past couple of years, the PAT guidance has slowly but consistently led to the change of an increasing number of pharmaceutical production processes (e.g. [START_REF] Sistare | Process Analytical Technology: an investment in process knowledge[END_REF]). In addition, the implementation of the basic ideas of the PAT guidance by the pharmaceutical industry have also resulted in increased focus on this area in other regulated sectors, such as the food industry. This paper is an attempt to initiate a broader discussion within the biochemical engineering community on how the PAT guidance influences the required skills of scientists, engineers and managers involved in pharmaceutical process development and manufacturing. This of course also includes the current and future generations of biochemical and chemical engineering students. First, a concise literature overview is provided, to illustrate what is available in the literature with respect to PAT. This is followed then by a critical discussion and an outlook to the future.

PAT in the literature

When taking a closer look at the literature, a substantial number of articles have been published already reporting PAT case studies or PAT-related work. The emphasis in many reported projects immediately following publication of the PAT guidance was on the analytical aspects: the papers either (i) report the development and implementation of on-line analysis techniques (e.g. [START_REF] Cogdill | Process Analytical Technology case study part I: Feasibility studies for quantitative near-infrared method development[END_REF][START_REF] Wikström | On-line content uniformity determination of tablets using low-resolution Raman spectroscopy[END_REF])especially spectroscopic methods -or (ii) report on the development and application of chemometric models to interpret the data generated by on-line analysis (e.g. [START_REF] Sulub | Content uniformity determination of pharmaceutical tablets using five near-infrared reflectance spectrometers: A process analytical technology (PAT) approach using robust multivariate calibration transfer algorithms[END_REF]). The focus on analytical aspects in the early applications of PAT can be understood from a process control point of view, since development of online process monitoring tools is essential to provide on-line information about the process which can subsequently be applied by implementing suitable control strategies. On the other hand, Maes and Van Liedekerke [START_REF] Maes | The need for a broader perspective if process analytical technology implementation is to be successful in the pharmaceutical sector[END_REF] point out that this focus on analytical aspects in many cases can be explained also by a misunderstanding of the PAT concept, i.e. companies think of PAT in terms of current validation and quality control processes, focusing on the sensor and what it can measure rather than on the wider quest for process understanding. So, it seems that closing the loop, i.e. using the on-line sensors installed in the frame of a PAT project within a control loop, is a major challenge for industry. One explanation for this could be that people working on PAT projects in general do not have sufficient skills to design, tune and implement control loops. The most essential point when developing PAT applications is the availability of sufficient process knowledge, i.e. one has to know how a change in an input will influence the process output, for example the product quality. A number of authors have recently suggested that mechanistic modeling -a skill that most engineers possess -is an excellent way to represent the process knowledge [START_REF] Brass | Application of modelling techniques for the improvement of industrial bioprocesses[END_REF][START_REF] Dassau | Combining six-sigma with integrated design and control for yield enhancement in bioprocessing[END_REF][START_REF] Sin | Matrix notation for efficient development of first-principles models within PAT applications: Integrated modeling of antibiotic production with Streptomyces coelicolor[END_REF][START_REF] Rathore | Modeling of biopharmaceutical processes. Part 1: Microbial and mammalian unit operations[END_REF][START_REF] Kaltenbrunner | Modeling of biopharmaceutical processes. Part 2: Process chromatography unit operation[END_REF].

Indeed, the differential equations representing one or several unit operations in a production process inherently represent the input-output dynamics, which is precisely the type of information needed to pinpoint the causes of excessive variation of product quality or to select suitable actuators to counteract undesired variations in product quality. To this end, the model can be supplemented by a modeling toolbox including uncertainty and sensitivity analysis to assess the statistical quality (read as reliability) of the simulated scenarios [START_REF] Sin | Good modelling practice (GMoP) for PAT applications: Propagation of input uncertainty and sensitivity analysis[END_REF].

Crystallization, a key operation in most pharmaceutical processes, is a good example to illustrate how process knowledge can be used. The increased understanding of the crystallization process has lead to the use of first-principles based approaches for crystallization process design [START_REF] Fujiwara | First-principles and direct design approaches for the control of pharmaceutical crystallization[END_REF], and first-principles based models are increasingly used in the design of advanced control strategies for crystallization processes [START_REF] Nagy | Modelling and control of combined cooling and antisolvent crystallization processes[END_REF].

Despite the fact that the use of mechanistic models for process knowledge representation increases steadily, it should be emphasized that data-driven modeling also plays a prominent role in the development of PAT applications. Chemometric models, for example, can be helpful tools in acquiring process [START_REF] Kourti | Process Analytical Technology beyond real-time analyzers: the role of multivariate analysis[END_REF]. For complex operations, for example [START_REF] Kiviharju | Modeling and simulation of Streptomyces peucetius var. caesius N47 cultivation and εrhodomycinone production with kinetic equations and neural networks[END_REF][START_REF] Jenzsch | Estimation of biomass concentrations in fermentation processes for recombinant protein production[END_REF], data-driven modeling is the only feasible alternative to represent input-output dynamics because our level of understanding is simply not sufficiently high to formulate mechanistic models. One of the key advantages of data-driven models is that they can be solved extremely fast, and therefore this type of models is preferred if predictions are to be available in real-time, for example in a process control context.

Last but not least, while on-line analysis is excellent and essential within a PAT application, it is not a goal in itself. On-line analysis methods should rather be perceived as useful tools. In a PAT context they are to be supplemented by continuous development of process understanding and by implementation of appropriate control strategies. Only then will PAT projects become a complete success.

Discussion

According to the PAT guidance, the following can be listed as PAT tools: [START_REF] Pollard | Biocatalysis for pharmaceutical intermediates: the future is now[END_REF] Multivariate tools for design, data acquisition and analysis; applications in an efficient way? Or in other words, how can we educate people to use the above-mentioned PAT tools? It is our thesis that education should not focus exclusively on the tools themselves -this too often appears to be the current approach (see above). Instead, learning should be based on detailed and realistic case studies, where the PAT tools are put to work to help create innovative manufacturing processes. Such an approach will help students to get a good feeling for the practical problems that will occur in any production process that needs to meet stringent product quality requirements, and in particular would assist in teaching problem-solving skills. Working with case studies should also appeal to professionals already working in the pharmaceutical industry, and would allow them to update their knowledge and skills such that they are better prepared for the task at hand: collecting relevant process knowledge and based on that, developing suitable control strategies to obtain consistent process performance and the requested high product quality.

We are convinced that process modeling and simulation will play an important role here. Indeed, educating students in process monitoring and process control would arguably be easier through the availability of a number of realistic and industrially relevant modeling case studies that are validated by a group of experts from academia and industry, and made available freely for download. Note that the idea of using process modeling and simulation in pharmaceutical process development and production is not new. Petrides et al. [START_REF] Petrides | Bioprocess simulation: an integrated approach to process development[END_REF] already demonstrated in 1989 how modeling and simulation can be applied to bioprocesses for where optimization could be most rewarding. In 1995, Lonza demonstrated how process modeling and simulation can be used to shorten development time in the production process of a pharmaceutical intermediate [START_REF] Rohner | Applications of modelling for bioprocess design and control in industrial production[END_REF]. Nevertheless, it would appear that the idea didn't completely catch up with the pharmaceutical industry in the nineties, probably because modeling often is considered as a timeconsuming task or perhaps due to the "perceived" reluctance of regulatory bodies to accept such manufacturing procedures as pointed out in the FDA guidance (http://www.fda.gov/cder/guidance/6419fnl.pdf.). The literature overview in the previous section of the paper demonstrates convincingly that there is a renewed interest in mechanistic modeling and simulation of production processes. [START_REF] Maes | The need for a broader perspective if process analytical technology implementation is to be successful in the pharmaceutical sector[END_REF] suggest that PAT project teams could learn from relevant experiences accumulated by other industries. To illustrate that the availability of the above-mentioned industrially relevant case studies is not an unrealistic dream, it is certainly worth mentioning a couple of initiatives that were taken in other research fields over the recent past to promote research in process control and process monitoring. In the chemical engineering field, the Tennessee-Eastman challenge was established in 1993 [START_REF] Downs | A plant-wide industrial process control problem[END_REF]. The Tennessee-Eastman challenge consists of a complete model of a chemical plant, including definition of control objectives, manipulated variables (including constraints), on-line measurements (ideal and non-ideal) and process operating constraints. In particular, the definition of the constraints and the fact that also non-ideal sensors on process optimization, process control (e.g. [START_REF] Mcavoy | Base control for the Tennessee Eastman problem[END_REF][START_REF] Ricker | Nonlinear model predictive control of the Tennessee Eastman challenge process[END_REF]) and process monitoring (e.g. [START_REF] Chiang | Fault diagnosis in chemical processes using Fisher discriminant analysis, discriminant partial least squares, and principal component analysis[END_REF][START_REF] Lin | Nonlinear dynamic principal component analysis for on-line process monitoring and diagnosis[END_REF][START_REF] Russell | Fault detection in industrial processes using canonical variate analysis and dynamic principal component analysis[END_REF]).

Maes and Van Liedekerke

Likewise in the field of environmental technology, the use of benchmark systems for assessment of process performance, control system evaluation and similar purposes is well established, especially for the evaluation of wastewater treatment plant control strategies [START_REF] Stare | Comparison of control strategies for nitrogen removal in an activated sludge process in terms of operating costs: A simulation study[END_REF][START_REF] Lee | New monitoring technique with an ICA algorithm in the wastewater treatment process[END_REF]. Originally started more than a decade ago, several benchmark plant lay-outs have now been developed, including a specific benchmark plant for testing of process monitoring algorithms [START_REF] Rosen | Towards a common benchmark for long-term process control and monitoring performance evaluation[END_REF] and a benchmark plant with a focus on plant-wide control (dynamic interaction between different plant units considered) [START_REF] Jeppsson | Benchmark Simulation Model No 2 -General protocol and exploratory case studies[END_REF]. Each wastewater treatment plant benchmark lay-out consists of a complete model representing a general treatment plant, an associated control system, a benchmarking procedure and a set of evaluation criteria. The inclusion of the benchmark wastewater treatment plant layout as a predefined software tool in several commercial simulator packages as well as in stand-alone Matlab, FORTRAN and C++ implementations has contributed considerably to their success. In addition, the simultaneous implementation of 12 different unit process models in several software packages resulted in some unexpected "by-products": it has for example led to a set of validated models (e.g. [START_REF] Rosen | Implementing ADM1 for plant-wide benchmark simulations in Matlab/Simulink[END_REF]) and modeling tools (e.g. [START_REF] Zaher | Transformers for interfacing anaerobic digestion models to pre-and post[END_REF]) which have become a standard in the wastewater treatment modeling community.

With these successful experiences in mind, we suggest to start a similar modeling benchmark initiative for pharmaceutical manufacturing systems -biotech-based as well as chemical-based -similar to the above-mentioned Tennessee-Eastman challenge [START_REF] Downs | A plant-wide industrial process control problem[END_REF] and the wastewater treatment benchmark plants [START_REF] Rosen | Towards a common benchmark for long-term process control and monitoring performance evaluation[END_REF][START_REF] Jeppsson | Benchmark Simulation Model No 2 -General protocol and exploratory case studies[END_REF]. The prime purpose of this pharmaceutical manufacturing system benchmark initiative would be to promote and facilitate the development and implementation of innovative PAT applications as well as the training of students at universities and staff involved in pharmaceutical production facilities. Specifically for the training of operators and engineers, one could for example use model simulations to illustrate the interaction between operating conditions in a crystallization unit (e.g. temperature, etc) and final product properties. Another example could be to demonstrate how the operating mode of a reaction unit (e.g. batch versus continuous process operation) influences the product yield as well as the choice of control and monitoring algorithms. The expert panel should then reach a consensus on the dynamic models to be used for the unit processes, the sensors and the actuators that are part of these case studies. These models would then have to be implemented and validated -ideally in different software packages -and following validation the models could then be made publicly available for download. These publicly available validated models would be similar for example to the model repositories in the systems biology field (e.g. http://www.biomodels.net/). The model implementation and validation would undoubtedly be one of the most timeconsuming parts of the simulation benchmark model development. Finally, the expert panel should decide on a number of reference simulation scenarios for each case study, such that the users of the simulation benchmark models have a reference point with respect to the process performance that is to be obtained. In fact, it could even be such that a competition could be organized where students at different universities could compete, for example to obtain the best process performance assuming that a limited number of on-line measurements can be introduced on the process.

Note that the definition of the case studies should be such that they allow a realistic simulation of manufacturing systems for pharmaceuticals, which includes 15 would be to apply for funding at the European level for setting up a network activity. Likewise similar organizations in the USA and Asia could also assist in developing the methodology and the tools. We would however strongly encourage industry (pharmaceutical production and software vendors) to be involved also,

i.e. it should not become a purely academic exercise.

Assuming that a benchmarking initiative for pharmaceutical manufacturing systems could be initiated, every participant in the initiative would clearly benefit.

For academia, students confronted with realistic simulation-based case studies would receive high-quality training into monitoring and control problems that are highly relevant for industry. Pharmaceutical production companies could employ graduated students with improved PAT-related skills, especially with respect to increased understanding of process dynamics and process monitoring/control. The benefit for the software vendors is that such a collaboration would create a forum where modeling case studies are validated such that they could be used to demonstrate simulation packages to a potential customer. For the research community, the participation of software vendors and pharmaceutical companies would have the additional benefit of promoting the dissemination of the work done in the framework of such a benchmarking initiative.

Conclusions and perspectives

Process Analytical Technology has changed pharmaceutical production, but has also created a need for the development of new skills for the scientists, engineers

  However, with the publication of the Process Analytical Technology (PAT) guidance in 2004 (http://www.fda.gov/cder/guidance/6419fnl.pdf.), the situation for the pharmaceutical manufacturing has changed significantly: One of the central issues in the FDA PAT guidance is indeed the transition from a fixed to a robust and adjustable manufacturing process by adopting innovative and new technology-based systems, as illustrated in the bottom part of Figure1. When implementing the PAT guidance on pharmaceuticals production, the process operation is adjusted as a consequence of upstream changes, such as changes in the properties of the raw materials. Instead of laborious off-line testing of product quality, the 'real-time release' concept is introduced in the PAT guidance, where product quality is now assessed in real-time based on the interpretation of on-line measurements. The focus of the PAT guidance is clearly on obtaining safe -for and efficient pharmaceutical manufacturing processes, but for industry the PAT guidance is also an open invitation towards development and implementation of innovative manufacturing processes. Indeed, as long as the critical process variables are kept within the "design space" of the process, it is possible to perform process optimization, for example by introducing more advanced control combined with on-line measurement of critical process variables.

  discussed in detail byKourti

  [START_REF] Johnson | Human insulin from recombinant DNA technology[END_REF] Process analysis equipment and tools (e.g. including software sensors); (3) Process control tools;(4) Continuous improvement and knowledge management tools (e.g. artificial intelligence, supervisory control). The question is now how we can provide the necessary education for the scientists and engineers involved in pharmaceutical process development and manufacturing to allow them to develop PAT

  economics and to identify precisely those process points

  a more realistic validation of process monitoring and control algorithms. The Tennessee-Eastman challenge -originally implemented in Fortran -has subsequently been implemented in other software packages (see e.g. http://depts.washington.edu/control/LARRY/TE/download.html), thereby extending the number of potential users considerably. As a consequence the Tennessee-Eastman challenge has been used extensively in research and teaching

Such a modeling benchmark

  initiative would require setting up an international expert panel. The expert panel should first agree on a number of suitable casestudies, for example one fermentation-based case-study and one based on organic

  in pharmaceutical manufacturing process development, for example within understanding, design, tuning and implementation of control algorithms. Based on experiences in other engineering fields, we conclude that mechanistic modeling applied to a number of relevant case studies, and supported by experts from academia and industry, could form the foundation for a benchmark modeling effort of pharmaceutical manufacturing systems. The result of such a benchmarking effort -documented and validated models of selected case-studies, endorsed by experts and publicly available for download -could significantly contribute to educating students and professionals with skills that are essential for implementing the Process Analytical Technology guidance in practice. a plant-wide modelling context. Environ.Model. Softw. 2007, 22, 40-58.

Figure 1 .

 1 Figure 1. Comparison between traditional 'fixed process' pharmaceutical

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Conflict of interest statement

The authors have declared no conflict of interest.


Today's biotech industry in India

Table 1 :

 1 Recent milestones of Indian biotechnology

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	11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37	1978 1981 1984 1986 1987 1989 1991 1996 1997 1998	The first Indian biotech company, Biocon, is established in Bangalore The Center for Cellular & Molecular Biology (CCMB) is founded in Hyderabad The Institute for Microbial Technology (IMTECH) is established in Chandigarh The Department of Biotechnology (DBT) is set up by the government of India to promote modern biology and biotechnology The National Institute of Immunology (NII) set up by DBT Bangalore Genei starts production of restriction enzymes & other tools for genetics The National Center for Biological Sciences (NCBS) is established to pursue R&D in molecular biology Bharat Biotech International Ltd. starts an R&D centre at Genome Valley, Hyderabad The Center for Biochemical Technology (CBT) is established to focus on bioinformatics and genomics Shantha Biotech launches India's first recombinant product, a Hep B vaccine Monsanto research establishes an R&D center for plant genomics
	38		DBT approves Monsanto to conduct trials with
	39		genetically modified cotton
	40		NCBS scientist sets up AVESTHAGEN, a plant
	41	1999	genomics company.
	42		The Indian states of Andhra Pradesh,
	43		Karnataka, Maharashtra and Tamil Nadu
	44	2000	announce biotech initiatives
	45		Reliance sets up Reliance Life Sciences to
	46		p0ursue stem cell based research and
	47	2001	product development
	48		Gujarat government announces Biotech
	49		Venture Capital Fund
	50		Wockhardt launched Wosulin (recombinant
	51		human insulin), Biovac -B (hepatitis B
	52	2003	vaccine) and Wepox (erythropoitin)
	53		Ocimum Biosolution entered into partnership
	54	2004	with US-based Genome Exploration
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Table 1 :

 1 India's top-20 home-grown pharmaceutical companies (left) and top-20 companies including global players (right). For more detailed data see supplementary information online

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	9 10 11 12	Rank 2008 1	Company Serum Institute of India	Revenues in $ Million 246.75	Rank 2008 1	Company Serum Institute of India	Revenues in $ Million 246.75
	13	2	Biocon	228.00 2	Biocon	228.00
	14 15	3	Panacea Biotec	169.50 3	Panacea Biotec	169.50
	16 17	4	Nuziveedu Seeds	75.75 4	Nuziveedu Seeds	75.75
	18 19 20 21 22	5 6 7	Rasi Seeds Indian Immunologicals Mahyco	73.32 5 49.00 6 42.50 7	Rasi Seeds NovoNordisk Novozymes South Asia	73.32 65.00 56.25
	23 24	8	Syngene International	40.00 8	Indian Immunologicals	49.00
	25	9	Jubilant	39.75 9	Mahyco	42.50
	26 27	10	Shantha Biotech	37.50 10	Syngene International	40.00
	28 29	11	Bharat Serums	35.00 11	Jubilant	39.75
	30 31	12	Bharat Biotech	31.50 12	Shantha Biotech 37.50
	32 33	13	Themis Medicare	27.50 13	Bharat Serums	35.00
	34 35	14	Haffkine Biopharma	22.15 14	EliLilly	34.25
	36 37	15	Rossari Biotech	20.50 15	Bharat Biotech	31.50
	38 39	16	Ankur Seeds	18.75 16	Themis Medicare	27.50
	40 41 42	17 18	Advanced Enzymes Ocimum Biosolutions	17.33 17 16.25 18	Aventis Haffkine Biopharma	26.25 22.15
	43 44 45 46 47	19 20	Nath Seeds Concord Biotech Total biotech revenue	15.50 19 15.00 20	Rossari Biotech GlaxoSmithKline 20.10 20.50
	48 49 50		accounted for by the 20 companies	1221.55
	51 52 53		Total biotech revenue in the country	2568.50
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Jiri Damborsky

Pennsylvania Stephen Benkovic

Uwe Bornscheuer

Janusz Bujnicki

Federico Gago

Philipp Holliger

Karl Hult

Dick Janssen

Karl-Erich Jaeger

Romas Kazlauskas

Stefan Lutz

Adrian Mulholland

Sven Panke

Gregory Petsko

Juergen Pleiss

Manfred Reetz

Brian Shoichet

Burckhard Seelig

Dan Tawfik

Nick Turner

Rebecca Wade

The third event of this conference series was held at the beautiful city of San Feliu de Guixols north of Barcelona in the hotel Eden Roc being directly located at the Mediterranean Sea. In contrast to the previous meeting held in Greifswald [1], it was this time organized under the umbrella of the European Science Foundation and the European Molecular Biology Organization as an ESF-EMBO symposium and Professor Jiri Damborsky (Brno, Czech Republic) was responsible for the scientific program. The meeting was attended by 149 participants from 30 countries with the majority coming from Europe. Notably, many young scientists joined the meeting and there was also a good mix between academia and industry.

The scientific program comprised of 24 invited lectures and 14 short talks subdivided into several sessions dealing with (i) fundamentals of enzymatic catalysis, (ii) computer modelling in protein design, (iii) bioinformatics in protein design and (iv) directed evolution (and engineering) of biocatalysts. Thus, the conference represented an excellent forum to exchange the latest achievements and theories on our current understanding of how enzymes function, how their relationship can be identified and interpreted, and how computational methods can be used to support these studies or even to tailor-design novel enzymes. Furthermore, it became obvious that directed (molecular) evolution has become a robust and state-of-the-art tool in protein engineering. It is impossible to cover all lectures in this report, but certain trends and highlights should be mentioned:

((quote))
Directed (molecular) evolution has become a robust and state-of-the-art tool in protein engineering A clear trend was the increasing importance of computer modeling and software tools in protein engineering. Daniela Grabs-Roethlisberger (University of Washington, Seattle, USA) and Dan Tawfik (Weizman Institute of Science, Rehovot, Israel) exemplified this with their outstanding work on the de novo computational design of an enzyme able to catalyze the Kemp elimination -a reaction for which no natural protein is known to be active. A completely different approach was presented by Burckhard Seelig (University of Minnesota, St. Paul, USA), who created a catalytically active protein from scratch using mRNA display.

Yet another alternative is the design of artificial metalloenzymes, which was demonstrated by Marc Creus (University of Neuchatel, Switzerland). Using avidins as protein hosts, the designed proteins can catalyze for instance enantioselective hydrogenation, allylic alkylation and sulfoxidation. In silico screening of the vast information available in public databases of protein sequences and structures is increasingly facilitated due to the development of software and advanced modeling tools. This was exemplified for instance by Adrian Mulholland (University of Bristol, Bristol, UK) for the application of QM/MM methods and Henk-Jan Joosten (Bio-Prodict, Wageningen, The Netherlands) for a database to identify key residues for mutagenesis. These developments were supported by many examples of various proteins, where the engineering of a given protein was substantially facilitated as computation significantly reduces the number of variants to be created and increased the probability to find the enzyme with desired properties. Furthermore, these methods allow for a much better understanding of the sequence-function relationship of proteins.

The second half of the symposium was much more focused on biocatalysis and hence the application of enzymes was in the foreground, but also novel methods for mutagenesis were presented. For instance, Manfred Reetz (MPI, Mülheim, Germany) showed that iterative saturation mutagenesis and CASTing enabled the rapid creation and identification of a highly enantioselective epoxide hydrolase (where previous directed evolution experiments using error-prone PCR led only to a modest increase in selectivity) and he could show that several routes exist, which can lead by step-by-step mutation to a useful enzyme. Aurelio Hidalgo (University of Madrid, Madrid, Spain) reported on a new method (OSCARR), which enables a facile cassette mutagenesis to modify a set of amino acids rather than single mutations and already led to the creation of an esterase with lipase-like chain-length profile. The flexibility of proteins was also demonstrated in the talk by Stefan Lutz (Emory University, Atlanta, USA), who performed circular permutation on lipase CAL-B and could show that even alternative Nand C-termini can result in a functionally active protein.

A strong link to organic synthesis was obvious in the lectures by Dick Janssen (University of Groningen, Netherlands) and Nick Turner (Manchester Interdisciplinary Biocentre, Manchester, UK). Janssen showed that halohydrin dehalogenase represent highly versatile biocatalysts in the synthesis of a broad set of enantiopure β-substituted secondary and tertiary alcohols and by far exceed the synthetic utility of epoxide hydrolases. Turner also introduced new enzymes useful for synthetic purposes such as amine oxidases, alcohol oxidases and ammonia lyases. From all lectures in this area it became obvious, that protein engineering is now state-of-the art technology to tailor-design biocatalysts for a given target.

Hence, the use of computer modeling for rational protein design and of directed evolution with high-throughput screening/selection tools appear to be generally used tools to solve synthetic problems.

Still, basic research to support our understanding on how proteins function has its place and is of high importance to solve issues in applied biocatalysis. This was exemplified for example in the lectures by Stephen Benkovic (Pennsylvania State University, University A. Warshel (University of Southern California, Los Angeles, USA) both using the 'bench-mark' protein dihydrofolate reductase (DHFR) as model enzyme. Gregory Petsko (Brandeis University, Waltham, USA) described general rules for identification of binding sites on protein surfaces using experimental studies (crystallographic analysis in the presence of small organic molecules) as well as theoretical calculations (theoretical solvent mapping and QM calculations of titration curves of ionisable groups).

Recent trends and a possible outlook on protein engineering was the focus of a plenary discussion chaired by Stephen Benkovic, Dick Janssen, Romas Kazlauskas, Dan Tawfik and myself with Jiri Damborsky as the moderator. The participants agreed that great progress has been made in terms of tools and data generation as such, but an in-depth understanding of the data is still lacking. A combination of computational and experimental data sets within one project is necessary to broaden our understanding of structure-function relationships. The general trend in protein engineering is a combination of directed evolution and rational design, for which experimentally the need for high-throughput screening methods is still important to identify interesting hits in large mutant libraries. The de novo design of enzymes from scratch is still far off as current designs do not match natural enzymes in their catalytic efficiency. This is clearly a 'higher order problem'. Also, how enzymes work in cells is not well understood. Hence, more research is needed for improved production of enzymes and new host organisms are needed to be developed for production of difficult-to-express proteins. It was also commonly accepted, that there is an increasing gap between large amounts of data and how to understand them. For instance, annotations of some data stored in genomic and proteomic databases is wrong, and thus the use of statistical analyses is becoming increasingly important. Another point which was intensively discussed was artificial life: one of the great challenges of synthetic biology is to design and construct the living cell and protein engineering may provide useful molecular tools for this difficult task. For protein engineering, the understanding and design of loops was considered highly important as loops are known to play an important role for binding of ligands to the active sites and are involved in the evolution of new substrate specificities. Thus, the development of novel methods for engineering of loops will promote construction of enzymes with novel substrate specificities or functions. Finally, the holy grail in protein engineering was identified as the prediction of the protein tertiary structure from the primary amino acid sequence.

((quote))

Cell and protein engineering may provide useful molecular tools for synthetic biology Overall, the 3rd symposium in this series was a great scientific meeting and an excellent opportunity to learn and discuss the latest developments in the rapidly developing field of protein engineering. I am sure that the participants are eagerly waiting for the next conference, hopefully taking place in 2010.

Uwe Bornscheuer (Greifswald University) E-mail: uwe.bornscheuer@uni-greifswald.de The meeting will bring together internationally renowned speakers who will review the current state of knowledge in the field of protein design and evolution for biocatalysis. The symposium will be the third event in a series held in Paris (2002) and Greifswald (2006). The main objective of the 2008 meeting will be to intensify discussion and collaboration between experimentalists with theoreticians, which is needed for development of novel biocatalysts by combining directed evolution with rational protein design. The meeting will be highly interdisciplinary through the participation of speakers with various backgrounds and research interests. The lectures will span the fields of mechanistic enzymology, biocatalysis, protein crystallography, computer simulations, phylogenetics, structural bioinformatics and synthetic biology. A substantial amount of time will be devoted to poster presentations and informal discussion to promote exchange of ideas and dissemination of information. Young scientists are encouraged to actively participate and a number of lectures will be reserved for their presentations based on originality and scientific quality of submitted abstracts.

1

 1 Meeting report: Protein Design and Evolution for BiocatalysisOctober 25-30, 2008, Sant Feliu de Guixols, Spain

Formatted

Formatted

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  In addition to the excellent lectures, the conference also featured almost 100 posters demonstrating highest quality science, which created a perfect platform for extensive discussions among the participants.On the leisure side, participants had the opportunity to visit the beautiful city Girona during a half day excursion. The meeting closed with an excellent and delicious conference dinner framed by a very enjoyable professional performance showing classical Spanish and gipsy dances.

  , Sant Feliu de Guixols (Costa Brava) Spain 25-30 October 2008 With support from

Wiley-VCH Biotechnology Journal

I Tour Generali, 15th Floor I Brussels I Belgium


--------------------------------------------------------------------------------------Another innovation from Biocon was the introduction of pre-filled syringes of two life saving drugs -GCSF (granulocyte-colony stimulating factor) and EPO (erythropoietin). These innovative products were introduced in mid-2008 for the first time in India. Biocon's pre-filled syringe device incorporates features that provide for simple and safe handing, a discrete appearance of the injector and a retractable needle that reduces the chances of injuries during injection. The company's stock price has been stagnant for quite some quarters now. Nearly 60% of its revenues come from exports of various biopharmaceutical products and Biocon was forced to book a lot of mark-to-market losses on account of its decision to make forward contracts in US $ at a much lower rate. However, Biocon is poised to grow in the years to come if the big bet on a slew of innovative products come off successfully. "We are gearing up to increase our top-line performance in biopharma. So the year ahead should see us delivering a better growth in terms of biopharma. We have grown by more than 100 percent in branded formulations sales compared to the previous year. We expect the growth to continue at this rate because the base is small. Biocon stands committed to build big brands of products like Insugen, BIOMAb-EGFR, and Statix. The company entered into a tie-up with Monsanto in 1999 to commercialize hybrid cotton with Bollgard technology. It was the second company in the Indian seed industry to do so with its hybrids and many more hybrids are in the pipeline to cover all cotton-growing zones. "Multinational companies are very strong on the technology front. Among Indian seed companies, a few companies are struggling to attain technological capabilities, but they are very weak in financial status when compared to multi-national companies. The interference in the pricing of quality value-added hybrid seeds by state governments is creating a negative impact on indigenous seed companies in their efforts," M. Ramasami points out. Rasi's ongoing projects include sourcing and evaluation of germplasm lines; breeding for new lines with earliness, erect leaves, orange flint and orange yellow semi-dent grain colors; development of new single cross, modified single cross and threeway cross hybrids for evaluation and identification of high-yield hybrid combinations; and maintenance breeding for parental purity. Rasi group also explores new trends in the industry like corporate and contract farming, which will provide a win-win solution to corporate farmer, spinner, ginner and the seed company. ------------------------------------------------------------------------------------------------------------------------- ----------------------------------------------------------------------------------------------------------- From its humble beginnings in a garage in a Bangalore suburb in 1978 as a manufacturer of industrial enzymes, Biocon has traversed a long way to emerge as one of the companies to watch out for in the next decade. Not just because its founder, Mrs Kiran Mazumdar-Shaw, is more famous and the most visible face of women entrepreneurs in India.

Because, in its new role as an innovative company, Biocon is betting big on developing an oral version insulin. And change the way diabetes is treated in India and the world. The phase 2 trials of oral insulin are currently on and regulatory approval for the imminent phase 3 trials will determine the shape of things to come for India's most watched biotechnology company.

Of course, Biocon has other businesses too. Realizing the importance of drug discovery services and clinical services, two of its subsidiaries---Syngene International and Clinigene International---are helping to make Biocon emerge as an integrated player with the ability to offer a range of drug discovery and clinical trials to global companies. "Biocon's operational performance remains on a growth track. At a time when many sectors face a severe slowdown, we believe that Biocon has a unique opportunity to leverage our affordable R&D base and forge partnerships that will deliver large contract research, manufacturing and co-development opportunities over the next 5 years," said Biocon Chairman and Managing Director, Mrs Kiran Mazumdar-Shaw. A recombinant insulin, INSUGEN, using Pichia pastoris expression, launched in 2004, and a monoclonal antibody-based head and neck cancer drug, BioMAB EFGR in 2007, are keeping the company's profile, along with its statin products custom-made for a number of big pharma companies.

Biocon's focus is on novel biologics, mainly monoclonal antibodies (Mabs), which have resulted in a robust and rapidly advancing pipeline. Some of its key programs include the development of Oral Insulin (IN -105) in the treatment of diabetes mellitus and a new MAb (T1h) targeted against CD6 in the treatment of rheumatoid arthritis (RA). Other programs that are expected to enter preclinical development include BVX-10 (a novel human anti -TNF) and BVX-20 (a novel human anti-CD20).

Biocon, in 2007, acquired 70 percent controlling stake in German Pharma company, AxiCorp Gmbh, for a consideration of €30 million. This acquisition helped Biocon to market and distribute a range of pharmaceuticals including generics, biosimilars, biologics and innovative pharmaceutical products in Germany and Europe.

Biocon's BVX-20 is a novel human monoclonal antibody that binds to CD20, a protein located on both normal and malignant B-cells. After binding, BVX -20 kills B-cells by recruiting the body's own immune system. BVX-20 is intended for use in the treatment of patients with relapsed or chemotherapy-resistant follicular B-cell NHL and CD20-positive diffuse large B-cell NHL in combination with chemotherapy. This monoclonal antibody was developed in collaboration with Vaccinex, a US company, using ActivMab technology. Proof of concept has been shown in animal studies and the molecule is expected to start clinical trials in 2009.

Another innovation from Biocon was the introduction of pre-filled syringes of two life saving drugs---GCSF (granulocytecolony stimulating factor) and EPO (erythropoietin). These innovative products were introduced in mid-2008 for the first time in India. Biocon's pre-filled syringe device incorporates features that provide for simple and safe handing, a discrete appearance of the injector and a retractable needle that reduces the chances of injuries during injection.

((Fig. 6 syringe))

Commenting on the launch of the pre-filled syringes, Mazumdar-Shaw explained, "The pharmaceutical industry has registered an increasing demand in the market segment of pre-filled syringe systems over the past ten years. These systems have the endorsement of healthcare professionals worldwide. There is a growing trend of injectable drugs moving into the pre-fill syringe as a presentation format. Worldwide, pre-filled syringe volumes have reached more than two billion units. The opportunity for Indian biotech companies in pre-filled syringes is immense as India is being viewed as a low-cost, high-quality manufacturing base. It has the largest number of USFDA approved plants outside the US." The company's stock price has been stagnant for quite some quarters now. Nearly 60 per cent of its revenues come from exports of various biopharmaceutical products and Biocon was forced to book a lot of mark-to-market losses on account of its decision to make forward contracts in US $ at a much lower rate. However, Biocon is poised to grow in the years to come if the big bet on a slew of innovative products come off successfully.

  Founded by India's leading breeder of race horses, Cyrus Poonawalla, in 1966 just to make some vaccines, Serum Institute of India in Pune has today emerged as one of the world's leading producers of pediatric vaccine. Serum is also one of the largest global producers of measles and DTP vaccines in the world. Every second child born in the world gets the first shot of vaccination with the products made in Serumpresence in the vaccine market and exports its products to 140 countries. Leading the race in MMR and hepatitis B vaccines in India, Serum Institute of India was also lauded for coming up with vaccine products in the country at affordable prices. ((Fig.2fermenter)) retained the crown for three consecutive years. In recent times, the company has achieved many milestones. In 2007, it announced the launch of its indigenously manufactured low-cost Haemophilus influenza type b (Hib) conjugate vaccine under the brand name of HibPRO in India. HibPRO has been priced at Rs 375 or just US $ 8 for a single dose vial. HibPRO is a vaccine with clinically proven efficacy and safety at an affordable price. According to Dr Cyrus Poonawalla, "HibPRO has undergone series of clinical trials to prove its efficacy and safety in comparison with commercially available Hib conjugate vaccines. With the launch of HibPRO, Serum can extend the benefits of pentavalent or quadravalent vaccines at a very affordable price. These benefits are mainly fewer numbers of injections, lesser pain to the child and protection against multiple diseases such as Hib, hepatitis B, diphtheria, pertussis and tetanus in one injection." ((Fig. 3 washing)) © CyberMedia Publications In the past, Serum's products were supplied to international health agencies like WHO, UNICEF, and PAHO (Pan American Health Organization), and also to over 140 countries across the globe. Its vaccines are used in the national immunization programs of several countries. The measles/rubella vaccine is used in catch-up and mop-up campaigns of several countries to control the incidence of measles and rubella. A novel Hib vaccine , was launched in March 2007. Serum Institute has a capacity to produce over 100 million doses of this vaccine. The company supplies this new-age vaccine to GAVI (Global Alliance for Vaccines and Immunization), PAHO and UNICEF. According to Dr Poonawalla, "Serum Institute is the only indigenous manufacturers of Hib vaccine in India today unlike other vaccine manufacturers who import the bulk. Hib vaccine is fully manufactured in India." The company supplies the Hib vaccine at a low price compared to those offered by other leading global the main objective of Serum Institute of India Research Foundation is to conduct research in the fields of medical sciences and in natural and applied sciences. . 4 vaccine)) © PhotoDiscIn 2006, Serum Institute of India has set up Serum BioPharma Park, India's first biotech Special Economic Zone (SEZ), and developments on this project are still in the process. The park is adjoining SEZ will allow Serum Institute of India to avail various tax benefits such as income tax and import duty on capital goods. This encouraged a lot of foreign companies to partner with Serum Institute of India to avail and share these benefits. The SEZ will be spread across 55 acres in Hadapsar, Pune. Serum Institute will have 5-6 units in the SEZ. The project is expected to be completed by 2010. "The SEZ, which has currently been allocated 55 acres of notified land in phase would, based on availabilities and opportunities, be gradually expanded to 250 acres. The SEZ will stimulate economic activities by

(

  (Fig.5kiran)) Dr Kiran Mazumdar-Shaw © CyberMedia PublicationsFrom its humble beginnings in a garage in a Bangalore suburb in 1978 as a manufacturer of industrial enzymes, Biocon has traversed a long way to emerge as one of the companies to watch out for in the next decade. Not just because its founder, Kiran Mazumdar-Shaw, is more famous and the most visible face of women entrepreneurs in India. Because, in its new role as an innovative company, Biocon is betting big on developing an oral version insulin. And change the way diabetes is treated in India and the world. The phase 2 trials of oral insulin are currently on and regulatory approval for the imminent phase 3 trials will determine the shape of things to come for India's most watched biotechnology company. Of course, Biocon has other businesses too. Realizing the importance of drug discovery services and clinical services, two of its subsidiaries -Syngene International and Clinigene International -are helping to make Biocon emerge as an integrated player with the ability to offer a range of drug discovery and clinical trials to global companies. "Biocon's operational performance remains on a growth track. At a time when many sectors face a severe slowdown, we believe that Biocon has a unique opportunity to leverage our affordable R&D base and forge partnerships that will deliver large

  the treatment of diabetes mellitus and a new mAb (T1h) targeted against CD6 in the treatment of rheumatoid arthritis (RA). Other programs that are expected to enter preclinical development include BVX-10 (a novel human anti -TNF) and BVX-20 (a novel human anti-BVX-20 is a novel human monoclonal antibody that binds to CD20, a protein located on both normal and malignant Bcells. After binding, BVX-20 kills B-cells by recruiting the body's own immune system. BVX-20 is intended for use in the treatment of patients with relapsed or chemotherapy-resistant follicular B-cell NHL and CD20-positive diffuse large B-cell NHL in combination with chemotherapy. This monoclonal antibody was developed in collaboration with Vaccinex, a US company, using ActivMab technology. The proof of concept has been shown in animal studies and the molecule is expected to start clinical trials in

(

  -filled syringe systems over the past ten years. These systems have the endorsement of healthcare professionals worldwide. There is a growing trend of injectable drugs moving into the pre-fill syringe as a presentation format. Worldwide, pre-filled syringe volumes have reached more than two billion units. The opportunity for Indian biotech companies in pre-filled syringes is immense as India is being viewed as a lowcost, high-quality manufacturing base. It has the largest number of USFDA approved plants outside the US."

  envisioned by M. Ramasami, and was initiated in the year 1973 at Attur in Salem district in the southern Indian state of Tamil Nadu. Rasi Seeds made rapid advances from seed production and supply ventures to a commendable position in research and development (R&D) because of the remarkable services of R. Krishnamurthy, renowned for his extra long staple superfine cotton variety Suvin and the popular Medium Staple variety LRA 5166. Having excelled in R&D in cotton seeds, it went on to expand into a multi-crop, multi-location quality research movement and proved beneficial to the farming community. The company aims to create genetic variability to suit various climatic zones with wider adaptability by producing hybrids of various staple lengths, high strength and a biotic stress tolerance in cotton. The R&D unit is recognized by the Department of Scientific and Industrial Research, Ministry of Science and Technology, Government of India, New Delhi. ((Fig. 10 plant)) Rasi seeds plant © CyberMedia Publications Rasi Seeds was the second largest seller of Bt cotton (genetically engineered insectresistant cotton) seeds in India in 2007. The company has a wellmaintained research farm of 140 acres, about 55 000 square feet contained green house, separate ginning facility for germplasm and other breeding lines, state-of-theart fiber testing lab (HVI), wellequipped seed testing laboratory, and a separate dehumidified refrigerated facility for germplasm and breeding lines. The company also has a state-of-the-art biotech facility for testing the presence of Bt gene. Rasi is the recipient of a national award for R&D from the Department of Scientific and Industrial Research, Ministry of Science and Technology, Government of India, New Delhi. It is the second company to obtain approval for Bt cotton. Rasi Seeds did extensive research for crop improvement incorporating latest technology in breeding techniques and is aiming for hybrids with earliness, high quality and drought tolerance. The RCH 2 cotton variant especially created history by its remarkable yield in farmers' fields. The company released fourteen elite Bt cotton hybrids for commercial cultivation with approval of Genetic Engineering Approval Committee (GEAC), Ministry of Environment and Forests, Government of India. With that, Rasi emerged first in the number of Bt cotton hybrids released in India. Also many more hybrids are in the pipeline to cover different segments in India. ((Fig. 11 botanist)) © Digital Vision The company, well known for its pioneering efforts in hybrid cotton, proved itself again by ushering in the Bt cotton technology.

  It was No1 in 2006. Deleted: rasi seeds ¶ Changing the face of India bioagri ¶ Deleted: Deleted: ,…sft Deleted: MNC Deleted: MNCs Deleted: pointed out Mr Deleted: , managing director, Rasi Seeds. Deleted: ¶ Deleted: has … Deleted: ; its popular RCH 2 Bt. RCH 2 is one among the very few Deleted: compared to the two big giants of Indian biotechnology, Biocon and Serum Institute. However, the company, founded by an electrical engineer, Varaprasad Reddy, in 1993 and named after his mother to produce India's first recombinant hepatitis B vaccine, has an iconic status in Indian biotechnology sector. For Reddy took up the challenge to prove to the world that an Indian company could make recombinant vaccines with world class quality. Today, the company supplies nearly half the global requirements of hepatitis B company conducts exploratory research in the fields of oncology, infectious diseases and platform technologies. Shanvac-B is the first Indian hepatitis B vaccine to be prequalified by WHO, Geneva, for supplying to UN agencies globally. Today, Shantha caters to major international markets including Asia-Pacific, Africa, CIS and Latin America, in addition to supra-nations like UNICEF and PAHO. ((Fig. 14 tank)) © CyberMedia Publications While attending a global vaccine seminar in California in 1993, I had decided to make Hepatitis B vaccine in India. I tried to get the technology but US companies sniggered at an Indian organization's ability to make quality Hepatitis B vaccine. I took up the challenge and founded the company with borrowed family funds. With help from scientists at the Center for Cellular and Molecular Biology (CCMB), Hyderabad, we made the vaccine in India," surmised Reddy about the iconic start up. , Hyderabad, we made the vaccine in India." When Shantha's Hepatitis B vaccine was introduce at onetenth the price of the imported vaccine, it shook up the market. Prices crashed. Five other Indian companies too introduced the receombinant Hepatitis B vaccine and today India has the cheapest vaccine in the world. Shantha is the company that most first-timer entrepreneurs in Indian biotech have looked up to since 1996. ((Fig. 15 hepatits)) ((wiki commons))Shantha has also got WHO prequalification for its combination vaccine of DPT+ Hepatitis B, Shantetra, enabling supplies to the UN Agencies. Shantha is currently bought the stake from Oman-based financial firms, which had invested in Shantha a few years ago.

Font

  An investment of Rs 1,200 crore( $ 250 million) by Serum has been made for the purpose. This investment has been divided in various phases. The phase I investment will be a minimum of Rs 500 crore ( $ 125 million) for Serum BioPharma Park and the developers would raise funds through internal accruals of the company as well as external borrowings from financial institutions.

(

  (quote)) "The opportunity for Indian biotech companies in pre-filled syringes is immense as India is being viewed as a low-cost, high-quality manufacturing base. It has the largest number of USFDA approved plants outside the US."

Table 1 :

 1 Business revenues of the Serum Institute in MilionUS- $, 2004US- $, -2008

http://www.seruminstitute.com A.2.2. Biocon: Aiming for innovation leadership

	now		focused	on	building	a	global	base	through
	new	drug	discoveries,
	developments, and services.

((Box))

Biocon's strength lies in reinventing. The group divested its industrial enzymes business a couple of years back and is

/www.shanthabiotech. com by Narayanan Suresh and Ch. Srinivas Rao, Editors, BioSpectrum, Cyber Media (India) Ltd, Bangalore 560 001, India E-mail: sureshn@cybermedia.co.in

						organizations such as the CCMB and the Indian Institute of	Deleted: like
						Science, among many other	Deleted: ,
						national	Indian	research
						institutions. A research tie-up with	Deleted: Bhabha Atomic Research
						the	International	Vaccine	Center, National Institute of
						Institute,	Korea,	for	the	Immunology, Indian Institute of
						development and transfer of	Chemical Biology, Jawaharlal Nehru
						technology for various vaccines	University, International Center for
						has complemented Shantha's	Genetic Engineering and
						research strengths.	Biotechnology, and National Center
						for Cell Sciences.
						Table 4: Business revenues of	Deleted: ¶
	This alliance favors synergies in terms of research and clinical development between the group companies. Shantha Biotechnics also has access to the support provided by Merieux Alliances international network. Shantha extends its excellent in-house expertise, core competences and skill in development of vaccines and biologicals to carry out contract Contract research is undertaken research services to its clients.	Formatted: Font: 8 pt pt Formatted Formatted: Font: 8 pt Formatted Deleted: F http:/Formatted: Font: (Default) Arial, 8 Shantha in Milion US-$, 2004 -2008 Deleted:
	at Shantha's sprawling 50 000	Formatted
	square feet R&D center. The
	center is well-equipped with the
	latest-world class instruments and other support facilities.	Formatted
	Shantha already carried out	Formatted: Caption
	substantial work for its clients.
	The contract research work
	undertaken is in the areas of
	gene	cloning,		monoclonal
	antibody			development,	Formatted
	expression and purification of
	recombinant		proteins	by
	fermentation, fermentation and
	purification process optimization,
	development	of	polyclonal	Formatted: Bullets and Numbering
	antibodies, and development of WCB, MCB and analytical	Formatted
	methods.
	((Fig. 16 scientists))
	© CyberMedia Publications
	Shantha	targets	infectious
	diseases and oncology as its	Formatted: Font: 8 pt
	primary areas of interest for conducting R&D activities. It also	Formatted
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						Formatted
	with	premier	national	and
	international			research

Profiles of four top biotechnology companies in India by Narayanan Suresh and Ch. Srinivas Rao

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			401, 4th Floor, MB Centre
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serum institute of india India's top BioPharma company

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- COMPANY STATS CEO: Dr CS Poonawalla (Chairman)

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	Business Revenue
	2004-05	Rs 505 crore ($112 million)
	2005-06	Rs 703 crore ($156 million)
	2006-07 2007-08 Page 2: [42] Formatted Rs 951 crore ($211 million) Rs 987 crore ($219 million) F o Font: (Default) Arial, 8 pt, Not Italic r Page 2: [43] Formatted	MvonElster MvonElster	2/12/2009 12:06:00 PM 2/12/2009 12:06:00 PM
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	Address: Tel.: Fax: Website: Page 2: [28] Formatted 212/2, Hadapsar, Font: (Default) Arial, 8 pt, Bold Page 2: [29] Formatted Body Text Page 2: [30] Formatted Font: (Default) Arial, 8 pt Page 2: [31] Formatted	Off	Soli MvonElster Poonawalla MvonElster MvonElster MvonElster R e v i Road, e w 2/12/2009 12:12:00 PM Pune -411028 91-20-26993900 91-20-26993921 2/11/2009 1:32:00 PM 2/12/2009 12:12:00 PM 2/12/2009 12:12:00 PM
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Rasi Seeds] aims to create genetic variability to suit various climatic zones with wider adaptability by producing hybrids of various staple lengths, high strength and a biotic stress tolerance in cotton."

  Number of columns: 3, Col #1 width: 120.4 pt, Col #1 spacing: 35.4 pt, Col #2 width: 120.4 pt, Col #2 spacing: 35.4 pt, Col #3 width: 120.4 pt, Not Force equal column width Bt for the central and south zones, RCH 118 Bt, RCH 138 Bt, RCH 144 Bt RCH 377 Bt for the central zone, RCH 134 Bt and RCH 317 Bt RCH 308 Bt and RCH 314 Bt for the north zone, and RCH 20 Bt and RCH 368 Bt RCH 371 Bt, RCH 111 Bt and RCHB 708 Bt for the south zone. Also

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	Page 2: [47] Formatted Font: (Default) Arial, 8 pt, Bold Page 2: [48] Change Formatted Bullets and Numbering Page 2: [49] Formatted Font: (Default) Arial, 8 pt Page 2: [50] Formatted Font: (Default) Arial, 8 pt Page 4: [63] Formatted Font: (Default) Arial, 8 pt, Not Italic Page 4: [64] Formatted Font: (Default) Arial, 8 pt Page 4: [65] Formatted Font: (Default) Arial, 8 pt Page 4: [66] Deleted Page 4: [66] Deleted Font: (Default) Arial, 8 pt, Bold Page 2: [51] Formatted Font: (Default) Arial, 8 pt it got approval for commercial release of its RCH 2 Page 4: [67] Deleted	MvonElster MvonElster MvonElster MvonElster MvonElster R e MvonElster v i MvonElster e MvonElster w 2/12/2009 12:06:00 PM 2/10/2009 12:33:00 PM 2/12/2009 12:06:00 PM 2/12/2009 12:06:00 PM 2/12/2009 12:06:00 PM 2/12/2009 12:06:00 PM 2/10/2009 12:32:00 PM MvonElster 2/6/2009 9:59:00 AM 2/12/2009 12:06:00 PM MvonElster 2/12/2009 12:06:00 PM MvonElster 2/10/2009 12:02:00 PM
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	rasi seeds Font: (Default) Arial, 8 pt Changing
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the face of India bioagri

  Rasi Seeds has been the first to engineer genetic variability to suit various agro climatic zones with wider adaptability by producing hybrids of various staple lengths, high strength and a biotic stress tolerance in cotton.

80 2852 3423 Website: www.biocon.com "Biocon stands committed to building big brands of products

  Bt. RCH 2 is one among the very few proprietary research hybrids that has been notified by the Government of India. The cotton hybrids RCH2, RCH 20, RCH 134 and RCH 138 were among the top ranking hybrids in the All-India Co-ordinated Cotton Improvement Project. We are gearing up to increase our top-line performance in biopharma. So the year ahead should see us delivering a better growth in terms of biopharma. We have grown by more than 100 percent in branded formulations sales compared to the previous year. We expect the growth to continue at this rate because the base is small. Biocon stands committed to build big brands of products like Insugen, BIOMAb-EGFR, and Statix."

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	F o r Page 4: [81] Deleted has done Page 4: [81] Deleted Page 5: [90] Change F Formatted Bullets and Numbering o Page 5: [91] Deleted has Page 4: [82] Formatted 2004-05 Rs 728 crore ($162 million) r 2005-06 Rs 793 crore ($176 million)	MvonElster MvonElster MvonElster MvonElster MvonElster	2/6/2009 5:36:00 PM 2/6/2009 5:36:00 PM 2/12/2009 12:11:00 PM 2/6/2009 10:16:00 AM 2/12/2009 12:06:00 PM
	Page 4: [72] Formatted Font: (Default) Arial, 8 pt Page 4: [73] Formatted Font: (Default) Arial, 8 pt, Not Bold Page 4: [73] Formatted Font: (Default) Arial, 8 pt Page 4: [74] Formatted P e MvonElster MvonElster MvonElster MvonElster e r R e v i e w 2/12/2009 12:06:00 PM 2/12/2009 12:06:00 PM 2/12/2009 12:06:00 PM 2/12/2009 12:06:00 PM Font: (Default) Arial, 8 pt Page 4: [83] Formatted MvonElster 2/10/2009 12:00:00 PM 2006-07 Rs 990 crore ($220 million) 2007-08 Rs 1090 crore ($242 million) P CEO: Dr. Kiran Mazumdar-Shaw (Chairman and Managing Director) Business: Manufacturer of biopharmaceuticals and industrial enzymes e Start-up Year: 1978 Address: 20th KM Hosur Road, e Electronic City, Bangalore -560100, India Tel.: +91 80 2808 2808 r R e v i e w Fax:+91 Page 5: [92] Formatted MvonElster 2/10/2009 12:01:00 PM Font: (Default) Arial, 8 pt, Not Bold Normal, Justified
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	, managing director, Rasi Seeds.
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" --Dr Kiran Mazumdar-Shaw, CMD, Biocon "Biocon's strength lies in reinventing. The group divested its industrial enzymes business a couple of years back and is now focused on building a global base through new drug discoveries, developments, and services.

Table 1 :

 1 Business revenues of the Serum Institute in MilionUS-$, 2004US-$, -2008

	Page 20 of 23

Table 1 :

 1 Business revenues of Biocon in MilionUS-$, 2004US-$, -2008

	Page 21 of 23

Table 1 :

 1 Business revenues of Rasi Seeds in MilionUS-$, 2004US-$, -2008

				Page 22 of 23
	80
	70
	60
	50
	40
	30
	20
	10
	0
	2004-05	2005-06	2006-07	2007-08

Table 1 :

 1 Business revenues of Shantha in MilionUS-$, 2004US-$, -2008

Page 23 of 23 Wiley-VCH Biotechnology Journal

Wiley-VCH Biotechnology Journal

Wiley-VCH Biotechnology Journal

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Wiley-VCH Biotechnology Journal