Presentation on theme: "QUALITY CONTROL OF SNAKE ANTIVENOM"— Presentation transcript:
1 QUALITY CONTROL OF SNAKE ANTIVENOM DR.M.V.KHADILKARTechnical Director,Premium Serums & Vaccines Pvt.Ltd.,Narayangaon,Dist.-PuneMaharashtra
2 PRODUCTION PROCESS Equine(Horse/Mules/Ponies) procurement Quarantine/Complete Health screeningRegular periodic Immunization with snake venomsPeriodic Titer TestingPlasma collection & storagePlasma FractionationBatch poolingSterile FiltrationContainerization & LyophilizationLabeling /InspectionFinished Product TestingMarketing
3 Antivenom quality and safety depends on 1.The control of source materials 2. Manufacturing procedures
4 Basic Raw Materials in Antivenom Production Snake VenomsHyperimmune Plasma
5 Objectives of the purification process High safety and efficacyNo impairment of the neutralizing activity during purificationMaintenance of high purityFree of microbial contaminants and endotoxinsWHO Workshop,
6 precautions in Venom production Venom is to be obtained from snakes in a controlled and documented manner. It should be properly preserved and tested for quality Venom being the prime raw material for the production of Antivenom ,it is a key factor in the production of effective antivenom.
7 Monitoring of immune response Proper collection process Plasma productionAnimal healthMonitoring of immune responseProper collection processare the greatest contributors to good quality plasma production
8 Collection of plasmaMaintenance of healthy & disease free herd of animalsCleanliness of premises and complete asepsis during procedure.Use of sterilized collection sets & containers under controlled environmental conditionsProperly designed clean room to separate plasma and operations carried out by trained personnel.Storage in proper temperature controlled rooms.Plasma collection vessels must be labeled with date, plasma volume, horse numbers and anti-coagulant use details.AVOIDS BIOBURDEN-HELPS IN CONTROLLING ENDOTOXINS
10 COMMON METHOD USED FOR PLASMA FRACTIONATION in India Pepsin Digestion to get F(ab’)2 fragments from whole IgG.Precipitation of unwanted proteins by either Ammonium Sulfate or Caprylic AcidConcentration & Diafiltration of F(ab’)2OTHER METHODSUse of whole IgG moleculePapain Digested Fab fragmentsChromatography (Ion-Exchange/Affinity)
11 PLASMA FRACTIONATIONUse of closed system with no exposure to environment.Least product hold time during processProcess validation-digestion, filtration, ultrafiltrationStrict supervision of critical process parameters such as Temp,pH,duration etcChecks about cleaning & sterilization statusIPQC checks -
12 Quality Control TESTS DESCRIPTION pH PROTEIN PRESERVATIVE CONTENT POTENCYABNORMAL TOXICITYSTERILITYENDOTOXIN(SOLUBILITY & MOISTURE CONTENT)
13 ANTIVENOM SAFETYDepends on 1. Type of antivenom -Whole IgG/ F(ab)2 / Fab Each fragment has different pharmacokinetics Efficacy reports not supported by clinical data 2. Dose 3. Route & Speed of administration 4. Manufacturing practices Hypersensitivity skin test has limited predictability value.
14 Reasons for reactions Contamination of plasma Enzymatic digestion & removal of unwanted proteinsFormulationSterilityNo data from Indian clinical trials is available, however , a lot has been reported by importing countriesThought to vary from 3-80% with about 5-10% severe reactionsReports are often subjective
15 EFFICACY MONOVALENT / POLYVALENT –Has its own advantages/disadvantages LEVEL OF REFINEMENT-Enzyme digestion,Ion exchange chromatography/Affinity chromatography. Effect on cost????VENOMQUALITYGEOGRAPHICAL ANTIGENIC VARIATIONSPECIFICITY/PARASPECIFICITYCLINICAL MANAGEMENT-OUTCOME DEPENDS ONEARLY START OF TREATMENTADEQUATE DOSESSUPPORT SYSTEMS
16 SNAKE ANTIVENOM I.P. Name of Product : Snake Antivenom I.P. (Polyvalent, Lyophilized/liquid, Equine origin Enzyme Refined Immunoglobulin)Description of Product :Lyophilized 20.0 ml VialAlong with 10.0 ml W.F.I. I.P. OR liquid 10 ml vialPotency : When Reconstituted With 10.0 mlof Sterile Water For Injection I.P.1.0 ml of reconstituted mixture Neutralizes0.60 mg. of Cobra Venom0.45 mg. of Krait venom0.60 mg. of Russell’s viper venom0.45 mg. of Saw scaled viper venomNature and concentrationof preservative : Phenol /Cresol not more than < 0.25%Storage conditions :Store in cool dark place, avoidexposure to sunlight for lyophilized and to store between 2-8˚C for liquid product.
17 How much antivenom is needed How much antivenom is needed?? ( Tumbare & Khadilkar,Journal of Bombay Veterinary college:2004,12(1&2):9-11)SnakeAverage Venom Yield in ml(Liquid)Average Venom Yield in mg (Lyophilized)Theoretical dose of antivenom required for treatment(10 ml vial)Indian Cobraml(0.435±0.01)mg(126.32±4.31)21Russell’s Viperml (0.412±0.02)mg(75.98±3.66)13Indian Common Kraitml(0.101±0.01)mg(8.35±0.64)2
18 INDIAN SCENARIOIt is estimated that, India currently produces about million Snake Antivenom vials per yearAlmost 75-80% of total Snake Antivenom produced is supplied to different government institutions via tender procedure.Current capacities can supply antivenom that meets about 60-70% of the requirements leading to unfortunate incidents of deathsMany times the problem is compounded by improper distributionThis results in big demand-supply gap for this life saving drug
19 INDIAN SCENARIOMany Indian companies have been regularly supplying this life saving medicine not only to its neighboring countries but also to countries in Africa and South Asia.Due to the enforcement of various regulations after 2000, manufacturing of equine biological products was significantly reduced resulting in severe shortage of these life saving products.Stringent regulatory requirements have also pushed up the production costs
20 INDIAN SCENARIOIndia still has very low price as compared to many other countriesIt has been observed that 2010 prices for a 10 ml vial of Indian polyvalent AVS range from about INR 300 to 500 (US$ 6.50–11.00), which is a fraction of the cost of a vial of CroFab antivenom in the USA (at over US$ 1900 per vial) or CSL antivenom in Australia (at US$ 1500 per vial)AVS is supplied by Indian antivenom producers to government hospitals at Rs 115 per vial (US$ 2.50)(Romulus Whitaker & Samir Whitaker in Current science, vol. 103, no. 6, 25 september 2012)Most of the ASVS tenders were awarded at Rs Rs. 210 per vial in 2011
21 My viewsDose finding studies can give guidelines for treatment protocol based on syndromic approach.One protocol for big country like India is difficult considering regional variation in venom composition.Use of polyvalent antivenom is more practical and economical especially considering procurement and distribution system followed.More legal sources of venoms should be established/promoted in different regions for its availability in production process.The Indian antivenoms are reasonably priced.There is limit to purification considering its direct effect on price.Collaboration between Industry & Academic institutions is needed to improve yields & reducing costs.