1. Company ProfilePanacea Biotecis Indias highly progressive, Innovative health management company based on Research & Development, Manufacturing and marketing of Pharmaceuticals, Biopharmaceuticals, Vaccines and natural / indigenous products.

The companyis witnessing a period of expansion across every aspect of our business from innovative products to customers in market, from manufacturing to regulatory approvals and thereby laying the foundation for translation of our vision in becoming greatest, largest and most admired biotechnology company in the World by 2020.

Ardent Research and Development efforts have always been a great strength ofthe company. The main research areas are New Chemical Entities (NCE), New Biological Entities (NBE) Novel Drug Delivery System (NDDS) based pharmaceutical formulations, Novel peptides & human monoclonal antibodies and Vaccine development. The company has developed four distinguished, ultra-modern, state-of-art R&D centers in different locations, having internal capabilities for constant research, with over 300 highly professional and skilled scientists engaged in various aspects of research.

Focused research efforts have led to grant of worldwide product patents valid in over 68 countries forPanacea Biotec. As on 31st March 2011, the company had filed over 1400 patent applications in various parts of the world including India. Of these, 382 have been granted patent and others are under various stages of examination or publication by the patent authorities. Some of these countries are USA, U.K., France, Germany, Italy, Sweden, Denmark, Spain, Finland, Switzerland, The Netherlands, New Zealand, Mexico, Brazil, Nigeria, Zimbabwe, Australia, South Africa, Japan, Russia, Canada, Ukraine, Korea and China,

The company has ultra-modern, state-of-art production facilities at Baddi (Himachal Pradesh), Larlu (Punjab) & Delhi for manufacturing tablets, capsules (including soft gelatin), ointments (transgel formulation) liquids, herbal formulations and vaccines. The facilities are WHO cGMP compliant.

The product portfolio includes highly innovative prescription products in niche therapeutic areas like pain management, diabetes & cardiovascular management, Oncology, renal disease management, osteoporosis management, anti-tubercular, gastro-intestinal care products and vaccines. Our current business stems from our leadership segments in India i.e. Nephrologicals, Anti Diabetes & Pain with some exclusive products based on patented Drug Delivery System.

Panacea Biotecis the largest Vaccine manufacturing Company in India and is well acknowledged by the UN Health Agencies in partnering the Polio eradication initiative with supplies of millions of doses of WHO Pre-qualified Polio vaccine. As a sequel to the completion of full range of Oral polio vaccines (tOPV, mOPV1, mOPV3 & bOPV)the companyhas introduced the next generation Inactivated Polio vaccine (eIPV) vide a collaboration with the Netherland Vaccine Institute. This vaccine has found extensive usage in India for more than 3 years now and is registered in Bangladesh. IPV is also in advanced stage of registration in 10 countries word wide with the target of being in more than 30 countries in a couple of years and has been put up for WHO prequalification which is expected soon.

Itis the first company in to have developed fully liquid Pentavalent vaccine (DTwP+Hep B+Hib) Easy Five. Easy Five is a WHO Prequalified vaccine being used in more than 25 countries worldwide. The vaccines portfolio also consists of Enivac-HB (Hepatitis B vaccine), Enivac-HB Safsy, Ecovac-4 (DTwP+Hep B), Easyfour (DTwP+Hib), which are also WHO prequalified. Vaccines in the offing are- Anthrax, Dengue, Japanese encephalitis and several others. A strong innovative vaccine pipeline is on the anvil which would pave the way for a consistent introduction of Next Gen Vaccines in future.

Panacea Biotecis the third largest biotechnology company (as per ABLE Survey, 2011), as well as among the top 50 pharmaceutical companies (as per ORG IMS March 2010) of India. To tap newer opportunities,the companyhas organized its formulation marketing into Six SBUs - PRO Care, Diacar Alpha, Diacar Delta, GROW Care, Onco Trust, and Critical Care, which enables it to respond to changes in the industry and marketplace. The vaccines are marketed through Chiron Panacea Vaccines, a 50:50 joint venture with Novartis Vaccines, U.KThe company has tie up with National Institute of Immunology, India for Japanese Encephalitis candidate vaccine, Biotech Consortium India Ltd. for the development, manufacture and marketing of Anthrax vaccine, worldwide and with National Institute of Health, USA, for use of a peptide based product for generation of hair follicles and hair growth.

Panacea Biotechas also collaborated with Netherlands Vaccine Institute for Inactivated Polio Vaccine; NRDC- India for Foot & Mouth Disease vaccine for veterinary use and Bio Farma-Indonesia for Measles vaccine.

2. Introduction2.1 VaccinesAvaccineis a biological preparation that improves immunity to a particular disease. A vaccine typically contains an agent that resembles a disease-causing microorganism, and is often made from weakened or killed forms of the microbe. The agent stimulates the body's immune system to recognize the agent as foreign, destroy it, and "remember" it, so that the immune system can more easily recognize and destroy any of these microorganisms that it later encounters.The term ''vaccine'' derives from Edward Jenner's 1796 use of the term ''cow pox'' (Latin ''variol vaccin'', adapted from the Latin ''vaccn-us'', from ''vacca'' cow), which, when administered to humans, provided them protection againstsmallpox.The immune system recognizesvaccineagents as foreign, destroys them, and 'remembers' them. When the virulent version of an agent comes along the body recognizes the protein coat on thevirus, and thus is prepared to respond, by (1) neutralizing the target agent before it can enter cells, and (2) by recognizing and destroying infected cells before that agent can multiply to vast numbers. Vaccines have contributed to the eradication ofsmallpox, one of the most contagious and deadly diseases known to man. Other diseases such asrubella, polio,measles,mumps, chickenpox, and typhoidare nowhere near as common as they were a hundred years ago.2.2 ImmunizationImmunization can be achieved in two stages- active or passive. In each case immunity can be acquired either by natural processes (usually by transfer from mother to fetus or by previous infection by the organism) or by artificial means such as injection of antibodies or vaccines.The types of immunization include:Passive ImmunizationAIM- Transient protection or alleviation of an existing conditionIn passive immunization, preformed antibodies are transferred to a recipient; in the past, passive immunization provided a major defense against various infectious diseases. Currently, passive immunization is used in several conditions like:i. Deficiency in synthesis of antibody.ii. When a susceptible person is exposed to a disease.iii. When sufficient immunization is not provided by active immunization.iv. When a disease is already present

Active ImmunizationAIM- To elicit protective immunity and immunogenic memoryIt can be achieved by natural infection with a microorganism or it can be acquired artificially by administration of a vaccine. After active immunization, a subsequent exposure to a pathogenic agent elicit`ts a heightened immune response that successfully eliminates the pathogen and prevents the disease.

2.3 Types of VaccinesAttenuated Viral or Bacterial vaccines - Microorganisms can be attenuated so that they lose their ability to cause significant disease but retain their capacity for transient growth within an inoculated host. Attenuation often can be achieved by growing a pathogenic bacterium or virus for prolonged periods under abnormal conditions.The ability of many attenuated vaccines to replicate within host cells makes them particularly suitable for inducing a cell-mediated response. Inactivated Viral or Bacterial vaccines - Another approach in vaccine production is to inactivate the pathogen by heat or by chemical means so that it is no longer capable of replication in the host. Chemical inactivation with formaldehyde or various alkylating agents has been successful.

2.3.1 Purified Macromolecules as Vaccines Polysaccharide vaccine: The virulence of some pathogenic bacteria depends primarily on the anti-phagocytic properties of their hydrophilic polysaccharide capsule. Coating of the capsule with antibodies and/or complement greatly increases the ability of macrophages and neutrophils to phagocytose such pathogens. These findings provide the rationale for vaccine consisting of purified capsular polysaccharide.

Toxoid Vaccine: Some bacterial pathogens, including those that cause diphtheria and tetanus, produce exotoxins. These exotoxins produce many of the disease symptoms that result from infection. Diphtheria and tetanus vaccines, for example, can be made by purifying the bacterial exotoxin and then inactivating the toxin with formaldehyde to form a toxoid. Vaccination with toxoid induces antitoxoid antibodies, which are also capable of binding to the toxin and neutralizing its effect. So formed toxoid are called formol toxoid.Recombinant Antigen Vaccines: The first recombinant antigen vaccine approved for human use is the Hepatitis B vaccine. This vaccine was developed by cloning the gene for hepatitis B virus (HBsAg) in yeast cells (Pichia pastoris). This recombinant Hepatitis B vaccine has been shown to induce the production of protective antibodies. 2.3.2 Recombinant-Vector Vaccines:It is possible to introduce genes that encode major antigens of especially virulent pathogens into attenuated viruses or bacteria. The attenuated organism serves as a vector, replicating within the host and expressing the gene products of the pathogen.DNA Vaccines: Plasmid DNA encoding antigenic proteins is injected directly into the muscle of recipient. Muscle cells take up the DNA and encoded protein is expressed, leading to both humoral antibody response and a cell-mediated response.

Synthetic Peptide Vaccines: Peptides are not as immunogenic as proteins and it is difficult to elicit both humoral and cellular immunity to them. The use of conjugates and adjuvants can assist in raising protective immunity to peptides, but barriers to the widespread use of peptide vaccines remain and pose an interesting problem for immunologists.

2.4 Some Processes associated with preparation of VaccinesAttenuationTo "attenuate" is to weaken a live micro-organism by ageing it or altering its growth conditions. This is accomplished by serial passage, that is, passing the live micro-organism through animal tissue several times to reduce its potency. For example, measles virus is passed through chick embryos, poliovirus through monkey kidneys, and the rubella virus through human diploid cells - the dissected organs of an aborted foetus.Vaccines made in this way are often the most successful vaccines, probably because they multiply in the body thereby causing a large immune response. However, these live, attenuated vaccines also carry the greatest risk because they can mutate back to the virulent form at any time.DetoxificationSome vaccines are made from toxins. In these cases, the toxin is often treated with aluminium or adsorbed onto aluminium salts to decrease the toxin's harmful effects. After the treatment, the toxin is called a "toxoid". Examples of toxoids are the diphtheria and the tetanus vaccines. Vaccines made from toxoids often induce low-level immune responses and are therefore sometimes administered with an "adjuvant", an agent that increases the immune response.

HEPATITIS-B VACCINEHepatitis - B vaccine (rDNA) is a non-infectious recombinant DNA Hepatitis B vaccine. It contains purified surface antigen of the virus obtained by culturing genetically-engineered Pichia pastoris yeast cells having the surface antigen gene of the Hepatitis B virus. The Hepatitis-B surface antigen (HBsAg) expressed in the cells of Pichia pastoris is purified through several chemical steps and formulated as a suspension of the antigen adsorbed on aluminium hydroxide and thiomersal is added as preservative.

2.5 Good Manufacturing PracticesGood manufacturing practice (GMP) (also referred to as cGMP or Current Good Manufacturing Practices)is that part of quality assurance which ensures that products are consistently produced and controlled to the quality standards appropriate to their intended use and as required by the marketing authorization. GMP is aimed primarily at diminishing the risks inherent in any pharmaceutical/biotechnological production, which may broadly be categorized in two groups: cross contamination/mix-ups and false labelling. Above all, manufacturers must not place patients at risk due to inadequate safety, quality or efficacy; for this reason, risk assessment has come to play an important role in WHO quality assurance guidelines.Agood manufacturing practice(GMP) is a production and testing practice that helps to ensure a quality product. Many countries have legislated that pharmaceutical and medical device companies must follow GMP procedures, and have created their own GMP guidelines that correspond with their legislation. Basic concepts of all of these guidelines remain more or less similar to the ultimate goals of safeguarding the health of the patient as well as producing good quality medicine, medical devices or active pharmaceutical products. In the U.S. a drug may be deemed adulterated if it passes all of the specifications tests but is found to be manufactured in a condition which violates current good manufacturing guidelines. Therefore, complying with GMP is a mandatory aspect in pharmaceutical manufacturing.Although there are a number of them, all guidelines follow a few basic principles: Manufacturing processes are clearly defined and controlled. All critical processes are validated to ensure consistency and compliance with specifications. Manufacturing processes are controlled, and any changes to the process are evaluated. Changes that have an impact on the quality of the drug are validated as necessary. Instructions and procedures are written in clear and unambiguous language. Operators are trained to carry out and document procedures. Records are made, manually or by instruments, during manufacture that demonstrate that all the steps required by the defined procedures and instructions were in fact taken and that the quantity and quality of the drug was as expected. Deviations are investigated and documented. Records of manufacture (including distribution) that enable the complete history of a batch to be traced are retained in a comprehensible and accessible form. The distribution of the drugs minimizes any risk to their quality. A system is available for recalling any batch of drug from sale or supply. Complaints about marketed drugs are examined, the causes of quality defects are investigated, and appropriate measures are taken with respect to the defective drugs and to prevent recurrence.GMP guidelines are not prescriptive instructions on how to manufacture products. They are a series of general principles that must be observed during manufacturing. When a company is setting up its quality program and manufacturing process, there may be many ways it can fulfill GMP requirements. It is the company's responsibility to determine the most effective and efficient quality process.

3. Clean RoomsA clean room is a controlled environment where products are manufactured. It is a room in which the concentration of airborne particles is controlled to specified limits. Eliminating sub-micron airborne contamination is really a process of control. These contaminants are generated by people, process, facilities and equipment. They must be continually removed from the air. The level to which these particles need to be removed depends upon the standards required.The only way to control contamination is to control the total environment. Air flow rates and direction, pressurization, temperature, humidity and specialized filtration all need to be tightly controlled. And the sources of these particles need to controlled or eliminated whenever possible. There is more to a clean room than air filters. Clean rooms are planned andmanufactured using strict protocol and methods. They are frequently found in electronics, pharmaceutical, biopharmaceutical, medical device industries and other critical manufacturing environments.

3.1 Clean Room Operating Parameters Room Cleanliness: Expressed as a classification per Fed. Std. 209E (e.g., Class 10, Class 100, Class 1,000, Class 10,000, Class 100,000), room cleanliness levels are the basis of any clean room design. It is important to know what classification you require, since an over-designed room can be costly to build and costly to operate. Room Temperature/Humidity: If the process is not affected by temperature/humidity conditions which fall outside of human comfort ranges, do not specify tight tolerances. A typical specification is 70 F2 and 30-70% RH. Tighter tolerance will result in unnecessarily escalated costs.Lighting Level: Level of 100 foot candles at the work surface is more than adequate for close assembly work. Higher lighting levels are costly in terms of energy consumption and initial cost. Lower levels can be maintained by control switching.Process Flow: This deals with the process being carried out within the room. The process will dictate the materials of construction as well as the layout of the facility. If the process is general in nature, a more flexible layout is recommended. The process should be defined as completely as possible before any clean room is laid out. This sounds elementary, but a poorly defined process will result in an unsatisfactory clean room, or one in which modifications are being made before the paint is dry. A knowledgeable clean room consultant will have experience in process flow and will be able to offer valuable assistance.

SOURCESISSUES

FacilityDust, bacteria, non-viable particulates

UtilitiesBacteria, non-viable particulates, hydrocarbons, aerosols

EquipmentBacteria, non-viable particles, hard to clean areas

ProcessNon-sterile connections, growth supporting process parameters, aerosols, metallic silvers, fibers, glass and rubber particles

PeopleViable and non-viable particles, skin cells, bacterial

4 .Key Elements of Contamination ControlHEPA (High Efficiency Particulate Air Filter)- These filters are extremely important for maintaining contamination control. They filter particles as small as 0.3 microns with a 99.97% minimum particle-collective efficiency.Clean Room Architecture- Clean rooms are designed to achieve and maintain a airflow in which essentially the entire body of air within a confined area moves with uniform velocity along parallel flow lines. This air flow is called laminar flow. The more restriction of air flow the more turbulence. Turbulence can cause particle movement.Filtration - In addition to the HEPA filters commonly used in cleanrooms, there are a number of other filtration mechanisms used to remove particles from gases and liquids. These filters are essential for providing effective contamination control.Clean Room Garments- Before entering the clean rooms a person should remove his street footwear and street clothes and wear cleanroom garments. The garments should be non-static and should shed minimum number of particles. Clean room garments also include Gloves, face masks and head covers

Limits For Particle Count:

Grades At Rest In Operation

Maximum no. of Particles Permitted per m3Maximum no. of Particles Permitted per m3

0.5-5m>5m0.5-5m>5m

A3500035000

B350003500002000

C350002000350000020000

D35000020000NDND

4.1Enviromental Monitoring ProgramAn appropriate microbial monitoring program should be established and used in production and laboratory facilities for both sterile and non-sterile products. The complexity of testing performed and the frequency of testing may vary depending upon the type of product, i.e. sterile versus non-sterile, the type of sterilization process, e.g., terminal sterilization versus aseptic filling and even within processes, e.g., overkill sterilization cycles versus combined biological indicator bioburden based cycles and absolute bioburden based cycles. Aseptic processing is the most stringent application of these principles.Limits for Microbial Contamination:

GRADESAIR SAMPLES cfu/m3SETTLE PLATES DIA 90cm/4hrsCONTACT PLATES, D55mm cfu/plateGLOVE PRINTS (5 FINGERS)

A