ASIA-PACIFIC BI TECHNEWS BIOTECHNOLOGY IN AN INTRODUCTION

D. Balasubramanian*

iotechnological activities in India can broadly be classified under two B categories — classical and modern. Classical biotechnology mainly refers to areas of biotechnology where tissues and organs, whole plants, organisms It is not commonly and animals are used, while modern biotechnology is largely based on molecular “ and cellular techniques. Classical biotechnology was already in practice effectively by a few universities and national agricultural research institutes in known that the areas India around the 1950s. Scholars trained in the classical disciplines of botany, zoology and agriculture were leading the research activities and indeed some of protoplast fusion, trail-blazing discoveries and developments were made in these universities. It is not commonly known that the areas of protoplast fusion, anther culture and callus culture, as well as micro-propagation using tissue culture of plants got anther culture and its earliest start in India, notably in the department of botany at the University of Delhi. The pioneers in these techniques are Professors Indra Vasil (protoplast callus culture, as well fusion), S. Maheshwari and his student Sipra Guha (callus culture and anther culture), B M Johri and H Y Mohan Ram (micropropagation, promiscuous as micro-propagation flowering of plants). Even as early as the late 1950s, micro-propagation of a variety of plants was successfully achieved in their laboratories. (It is thus notable, but not surprising, that much of the biotechnological activities in using tissue culture of India at present is still pretty much plant-based and is focused on micro- propagation in green houses.) It was around that time that plant geneticists plants got its earliest realized the importance of hybrid seeds, cross varieties, selection breeding and other biotechnological inputs. The Indian Council of Agricultural Research start in India. (ICAR), which predates independent India, had established excellent field stations and research outlets at various places such as Pusa, New Delhi, and Coimbatore. ”

*Professor D. Balasubramanian is Director of Research, Hyderabad Eye Research Foundation, L.V. Prasad Eye Institute, Hyderabad, India.

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BTV4•4/part1 47 2/9/00, 9:09 AM “The fact that a nation that was forced to import food two generations ago now produces more than its needs and is able to export food grains is testimony to the successful use of biotechnology in agriculture.

Concurrent with these national efforts in food production” Classical Biotechnology and livestock improvement, vaccination and other When the Mexican grass variety of wheat was introduced intervention methods have also been in practice on a large to the world by Norman Borlaug, one of the earliest scale in order to eliminate debilitating and death-causing countries to take advantage of this was India. Based on a diseases such as cholera, smallpox, measles, and goiter, systems approach, involving hybrid seed selection and among others. Delivery of the vaccines and immunization breeding, irrigation, use of appropriate fertilizers, post- of several hundred million people across a nation of harvest technologies and distribution of the produce, led 3000 km x 2000 km is an exercise of continental to what is now recognized as the Green Revolution. From proportions, which has been carried out successfully a mere 50 million tons output of food grains in the late more than once. The most recent instance has been the 1940s, India now produces 200 million tons — in just a immunization of over 150 million Indian children against matter of slightly more than 50 years. The fact that a nation polio using the oral polio vaccine in a ‘pulsed’ fashion that was forced to import food two generations ago now twice within 60 days. Similarly, the large-scale elimination produces more than its needs and is able to export food of goiter from focal areas through the use of iodized salt grains is testimony to the successful use of biotechnology and the administration of vitamin A doses to school children in agriculture. Amongst the active promoters of this form to counter night blindness have also been a story of of biotechnology are the scientists M S Swaminathan, success in India’s march towards better health. Drs. V Gurdev Khush, and M V Rao. Ramalingaswami and C Gopalan, two former chiefs of the Indian Council of Medical Research, have played key Classical biotechnology has also been used in India to roles in promoting and coordinating these efforts on a improve the livestock industry. Selective breeding of chosen national scale. breeds of milch cattle, sheep, horses and other animals has been successful. The White Revolution in dairy has closely Birth of Modern Biotechnology followed the Green Revolution in food production. Modern biotechnology was introduced in India around Cooperative dairy management involving thousands of the 1970s when a significant number of Indian researchers dairies, proper choice of livestock and milch cattle, an trained in the US and Europe in various aspects of modern efficient people-friendly, and people-participatory biotechnology returned to India. In fact, some of them distribution system led to this revolution. Varghese Kurien were front ranking players in the development of numerous is highly regarded in this matter. It is interesting to note methods and significant discoveries in enzymology, that while M S Swaminathan, Gurdev Khush and M V biopolymer structure and conformation, reverse transcrip- Rao are trained scientists, Kurien is a generalist and a tion, molecular virology, ribosome structure, transcription systems manager. This underscores another facet of and translation, cell culture techniques, biomolecular biotechnology, namely the close and necessary interaction spectroscopy, computer methods and so on. Indeed in the between the researcher and the manager. field of biopolymer conformation, the pioneering work of

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The DBT operates with an overseas scientific advisory committee, as well as an internal scientific advisory committee comprising of experts from within the country. The DBT not only funds extramural research projects but has also started institutions and centers of excellence in areas of modern biotechnology such as the National Institute of Immunology, National Centre for Cell Sciences, Centre for DNA Fingerprinting and Diagnostics, and the National Centre for Brain Research. It also supports university centers of excellence. The DBT has also prepared guidelines on the use of recombinant DNA techniques, transgenics, and other issues.

An important fillip to capacity building in biotechnology in the country has been provided by the DBT through their MSc programs in biotechnology, for which students are chosen on the basis of a national examination, and the DBT postdoctoral research program which supports about a hundred postdoctoral research fellows in four or five centers across the nation. Sponsored projects and grant-in- aid projects also employ postdoctoral researchers. International cooperation in the area of biotechnology has also been possible through the DBT Visiting Associateships and Visiting Fellowships offered to both Indian and overseas scholars.

G N Ramachandran had already established him as a world In addition to the DBT, other governmental agencies leader. His school in Madras and later in , such as the Indian Council of Medical Research (ICMR), comprising researchers such as V Sasisekaran (nucleic Indian Council of Agricultural Research (ICAR), acid conformations), V S R Rao (polysaccharides), Department of Science and Technology, Council of C Ramakrishnan (protein structure), M Vijayan (X-ray Scientific and Industrial Research (CSIR), and the Bureau crystallography), and their students and associates continues of Research in Nuclear Sciences (BRNS), as well as the to lead in the field of conformational predicts and analysis. Universities Grants Commission are other sources of It was about the same time that the Department of Science research funding in biotechnology in the country. It has and Technology (DST) of the Indian Government initiated been estimated that close to 60% of all grants-in-aid offered the National Biotechnology Board or NBTB. This board, by these national agencies goes into areas of modern through its two pioneering researchers — the bio-organic biology. Professional associations in the area of modern chemist S Varadarajan (then secretary of the DST) and the biology have also been active for over two decades now. biologist S Ramachandran (then head of the NBTB), played a proactive role in promoting specific methods and Notable amongst these are the Society of Biological technologies, curricular programs, research laboratories and Chemists of India, Indian Cell Biology Association, Indian funding research projects to individual scientists. Within a Immunological Society, Association of Indian Microbio- short time, NBTB matured into a full-fledged department logists, Indian Biophysics Society, and the informal and of the government, called the Department of Biotechnology yet very effective annual meeting called the Guha Research (DBT). During the 15 years of its existence, it has been Conference. These have led to camaraderie and cooperation led by prominent scientists such as S Ramachandran, C R between investigators within the country so that exchange Bhatia and currently, Manju Sharma, who have started of scholars, exchange of materials and sharing of equipment and sustained programs of education, training, research has become far more common in the areas of biotechnology and development in various areas of biotechnology in the and modern biology than in other branches of science in country. India.

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BTV4•4/part1 49 2/9/00, 9:10 AM Biotech products through basic research — Launch of leprosy vaccine (Leprovac); The vaccine was developed at the National lnstitute of Immunology and transferred for commercial production to Cadila Pharmaceuticals.

Contemporary Centers of Biotechnological Activity goes on in the ICAR laboratories and provincial agricultural Amongst the internationally known players in biotech- universities such as the ones in Hyderabad, Coimbatore nology are the Indian Institute of Science in Bangalore and Pantnagar. (a postgraduate institution of 80 years standing, with close Some of the more notable contributions in the area of to 85 faculty members in all areas of biology), the Tata modern biotechnology in India would be macromolecular Institute of Fundamental Research in Mumbai, and its conformation (theory and experiment), glycobiology and offspring the National Centre for Biological Sciences in lectin structure, procaryotic transcription and the mode of Bangalore, the Jawaharlal Nehru University in New Delhi, action of drugs such as rifampicin, choloroquine and so the Madurai Kamaraj University in Madurai, the Pune on, enzymological approaches to malaria control and University in Pune, the University of Hyderabad in drug design, molecular biology of cytochrome P450, Hyderabad, the M S University in Baroda, the Banaras peptide design, molecular mimicry, epitope-based vaccine Hindu University in Varanasi, and the Bose Institute in design, expression vectors for protein production, structural Calcutta. Equally important are the national laboratories basis of chaperone action, drug delivery systems, DNA such as the Centre for Cellular and Molecular Biology and typing and lineage analysis, and so on. the Centre for DNA Fingerprinting and Diagnostics, both in Hyderabad, the Indian Institute of Chemical Biology in Biotech Products through Basic Research Calcutta, the National Institute of Immunology in New A few notable biotech products have come through active Delhi, the Centre for Biochemical Technology in Delhi, industry-academic collaboration in the last few years. For the Central Drug Research Institute in Lucknow, the example, two medium range companies have been able to Institute of Microbial Technology in Chandigarh, and the produce and market effective hepatitis B vaccine, through UNDP — aided International Centre for Genetic collaborations with academic institutions. The mode of Engineering and Biotechnology in New Delhi (which has collaboration here is of interest. One of these companies its twin in Trieste, Italy). Significant agri-biotech research was actually housed at a national laboratory, with its

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scientists working as guest researchers, when the products were developed and validated. Once this was done, clinical trials, up-scaling and production followed. The company scientists enjoyed all the privileges of scientists from the academic institution — used the facilities of the host research laboratory and learnt the grammar of basic research, while the scientists from the institution had the feel for what it is like to meet deadlines, demands on research products and so on. It was what is called a win-win situation for both parties. The research laboratory profited in terms of revenue while the company had a host of talented scientists working as surrogates. Other products hitting the market through such collaborations are streptokinase, interferon, and a general- flourishing area where the major players are Mahyco, Indo- purpose salt inducible expression vector system, which is American Hybrid Seeds, SPIC, EID Parry, and lately of great use for both academic and large-scale production Monsanto. of proteins from recombinant DNA technology. This vector Among the major Indian drug firms are Ranbaxy, has been patented in the US Dr. Reddy’s Laboratories, Cadila, Unichem, and Cipla, and licensed to a company among others. The notable point about these companies there for global marketing. is that they are completely home grown and at present In addition, there is a whole have full fledged R&D laboratories associated with them. range of serum diagnostic Apart from classical methods of drug discovery, some of kits that have been developed them have dived right into the quest for new molecules. in academic institutions Three new molecules have already appeared on the drug and handed to the industry scene and are being licensed to firms abroad. The firm for marketing and sales. Biocon, located near Bangalore, is a success story in the Amongst the more notable of custom-production of enzymes and biochemicals for these is a diagnostic kit that specific clients. Astra Zeneca has its research and exploits an agglutination development center in Bangalore, where it concentrates on method using whole blood to the development of drugs for infective diseases, and high detect the presence of the AIDS virus. Laboratories in throughput screening of candidate molecules. Traditional academic institutions are developing cholera vaccine, Indian medicine and natural products is also an active area leprosy vaccine, and vaccine against the Japanese of research. Companies such as Dabur, Arya Vaidyasala encephalitis virus. While the leprosy vaccine has already Kottakal, among others, are focusing their attention on been licensed to a firm, the other two are expected to be looking for validation, promotion and marketing of drugs licensed shortly.

Turning to the industry, perhaps the most prevalent biotechnology industry in India in almost every major city “Traditional Indian medicine and natural products is that of tissue culture and micro-propagation of plants, is also an active area of research. shrubs and flowers. There are several major players such as SPIC, A V Thomas, Godrej, but there are also many A noteworthy national effort is the inter-laboratory small scale micro-propagators. It is interesting to note program of the CSIR to test and validate a large that during the severe winter of 1994–95 in Europe, tulips, number of bioactive natural products through the the favorite of the Dutch people, were exported in cargo use of chemical, biochemical and pharmacological loads by air from Hyderabad to Amsterdam — a methods of screening. biotechnological twist to the phrase ‘carrying coal to Newcastle’! Hybrid seed production and sales is another ”

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BTV4•4/part1 51 2/9/00, 9:10 AM based on traditional natural product. A noteworthy national Diagnostics (CDFD), National Institute of Nutrition (NIN), effort is the inter-laboratory program of the CSIR to test Dr. Reddy Research Foundation (DRF), Hyderabad Eye and validate a large number of bioactive natural products Research Foundation (HERF), International Centre for through the use of chemical, biochemical and pharmaco- Research in Semiarid Tropics (ICRISAT, largely devoted logical methods of screening. The CSIR has entered into to five coarse grain cereals), and the Ranga Agricultural a joint venture with Kottakal for certification of the University, Directorate of Rice Research (under Prof. E A Ayurvedic pharmacopia. Proteins, sugars, gums and resins, Siddiq who has developed successful rice hybrids), and enzymes from natural and selectively bred microbial University of Hyderabad (which collaborates with the sources are other aspects of the Indian biotechnology ICRISAT and DRF), Osmania University (with strength in industry that have done well. Today there are a couple medical and cytogenetics), Indian Institute of Chemical of local suppliers, such as Bangalore Genei (started by Technology (focused on bio-organic chemistry, bioactive Dr. P Babu, a former academic), which supply their compounds screening) and the Deccan and Gandhi Medical molecular biologicals to academic institutions, while Colleges. State-of-the-art medical centers and hospitals not competing with companies such as Sigma, Gibco-BRL, only provide medical services but also interact with research and Amhersham. Most of these local companies supply laboratories in the city. enzymes, oligos and probes, offer synthesis and sequencing In addition, support from the state government of Andhra services for both peptides and nucleic acids, and also loan Pradesh (of which Hyderabad is the capital city) in business small laboratory equipment to the academic institutions. development, particularly in the emerging areas of Hyderabad – the Biotech Capital of India information technology and biotechnology, has been full and proactive. A state-sponsored Biotechnology Park along The city of Hyderabad in South Central India has fast the lines of those already established in Singapore and turned out to be the hub of biotechnological activity. It is Hong Kong, is coming up, next to the already established here that the drug company, Dr. Reddy’s Laboratories ‘Knowledge Park’ in the suburbs of the city. The park (DRL), has its research lab and drug production center, provides all infrastructures — water, power, building and from where two new molecules have recently emerged – lab space, communication facilities, conference and seminar one of them licensed to Novo Nordisk. This demonstrates halls, customs and license clearance and other ‘house- the fact that the process of inventing a drug, testing it and keeping’ facilities. A few biotech firms have already putting it out in the market can be done more cost- negotiated for occupation in the Biotech Park, which is effectively from India and South Asia. It is in Hyderabad situated right next to the firm Bharat Biotech. that another notable pharmaceutical firm — Biological E Limited is located; and it is from here that two Until about a decade ago India’s economic policy was small firms — Shanta Biotechics and Bharat Biotech — pretty much governed and restricted by the government. developed vaccines against hepatitis B and marketed them In 1991, when globalization of the Indian economy was competitively and successfully. (These efforts also illustrate initiated, various restrictive practices and procedures were the effective and mutually enhancing collaboration between abandoned, and more and more private enterprise and joint industry and basic research laboratories — Shanta venture enterprises have been encouraged. This has led to Biotechics with the University of Missouri and with the a significant number of pharmaceuticals, agricultural, Centre for Cellular and Molecular Biology (CCMB) in cosmetic and biological firms from abroad entering the Hyderabad, and Bharat Biotech with the Indian Institute Indian scene. Some of these function as regional offices, of Science in Bangalore.) There are about half a dozen while others work in collaboration as joint ventures with companies around the city, drug formulating franchisees Indian entrepreneurs. The most recent entrant has been that work with global companies such as Glaxo, Procter Monsanto, which has also established its R&D center in and Gamble, and Astra Zeneca, and public sector Bangalore. While the rate of growth of the national (government-owned) drug companies such as the Indian economy has been a healthy 5% to 7% per annum, growth Drugs and Pharmaceuticals Limited and the Indian in biotechnology and medical sectors has been even higher. Immunologicals, which specializes in veterinary products. With pro-active policies, one can expect this figure to keep growing and India will emerge as one of the major players Hyderabad has some of the more notable research in both classical and modern biotechnology. The All India institutions such as the CCMB (basic and in part applied Biotechnology Association has been attempting to liaise research in biology), Centre for DNA Fingerprinting and with its international partners elsewhere towards this goal.

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