QUALITY CONTROL OF SNAKE ANTIVENOM DR.M.V.KHADILKAR Technical Director, Premium Serums & Vaccines Pvt.Ltd., Narayangaon,Dist.-Pune Maharashtra
Dec 16, 2015
QUALITY CONTROL OF SNAKE ANTIVENOM
DR.M.V.KHADILKAR
Technical Director,
Premium Serums & Vaccines Pvt.Ltd.,Narayangaon,Dist.-Pune
Maharashtra
PRODUCTION PROCESS Equine(Horse/Mules/Ponies) procurement Quarantine/Complete Health screening Regular periodic Immunization with snake
venoms Periodic Titer Testing Plasma collection & storage Plasma Fractionation Batch pooling Sterile Filtration Containerization & Lyophilization Labeling /Inspection Finished Product Testing Marketing
ANTIVENOM QUALITY AND SAFETY DEPENDS ON
1.The control of source materials
2. Manufacturing procedures
BASIC RAW MATERIALS IN ANTIVENOM PRODUCTION
Snake Venoms
Hyperimmune Plasma
OBJECTIVES OF THE PURIFICATION PROCESS
High safety and efficacy
No impairment of the neutralizing activity during purification
Maintenance of high purityFree of microbial contaminants and
endotoxins WHO
Workshop,
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.
PLASMA PRODUCTION
Animal health
Monitoring of immune response
Proper collection process
are the greatest contributors to good quality plasma production
COLLECTION OF PLASMA Maintenance of healthy & disease free herd of
animals Cleanliness of premises and complete asepsis
during procedure. Use of sterilized collection sets & containers
under controlled environmental conditions Properly 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
ENZYMATIC CLEAVAGE OF IMMUNOGLOBULIN MOLECULE
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 Acid
Concentration & Diafiltration of F(ab’)2
OTHER METHODS Use of whole IgG moleculePapain Digested Fab fragmentsChromatography (Ion-Exchange/Affinity)
PLASMA FRACTIONATION
Use of closed system with no exposure to environment.
Least product hold time during process Process validation-digestion, filtration,
ultrafiltration Strict supervision of critical process parameters
such as Temp,pH,duration etc Checks about cleaning & sterilization status IPQC checks -
QUALITY CONTROL TESTS
DESCRIPTION pH PROTEIN PRESERVATIVE CONTENT POTENCY ABNORMAL TOXICITY STERILITY ENDOTOXIN (SOLUBILITY & MOISTURE CONTENT)
ANTIVENOM SAFETYDepends on
1. Type of antivenom -Whole IgG/ F(ab)2 / FabEach fragment has different pharmacokineticsEfficacy reports not supported by clinical data
2. Dose3. Route & Speed of administration 4. Manufacturing practices
Hypersensitivity skin test has limited predictability value.
REASONS FOR REACTIONS
Contamination of plasma Enzymatic digestion & removal of unwanted
proteins Formulation Sterility
No data from Indian clinical trials is available, however , a lot has been reported by importing countries
Thought to vary from 3-80% with about 5-10% severe reactions
Reports are often subjective
EFFICACY
MONOVALENT / POLYVALENT –Has its own advantages/disadvantages
LEVEL OF REFINEMENT-Enzyme digestion,Ion exchange chromatography/Affinity chromatography. Effect on cost????
VENOM QUALITY GEOGRAPHICAL ANTIGENIC VARIATION SPECIFICITY/PARASPECIFICITY
CLINICAL MANAGEMENT-OUTCOME DEPENDS ON EARLY START OF TREATMENT ADEQUATE DOSES SUPPORT SYSTEMS
SNAKE ANTIVENOM I.P.
Name of Product Name of Product : : Snake Antivenom I.P.Snake Antivenom I.P. (Polyvalent, Lyophilized/liquid, Equine origin (Polyvalent, Lyophilized/liquid, Equine origin
Enzyme Refined Immunoglobulin)Enzyme Refined Immunoglobulin) Description of ProductDescription of Product :Lyophilized 20.0 ml Vial :Lyophilized 20.0 ml Vial Along with 10.0 ml W.F.I. I.P. OR Along with 10.0 ml W.F.I. I.P. OR
liquid 10 ml vialliquid 10 ml vial Potency Potency : When Reconstituted With 10.0 ml : When Reconstituted With 10.0 ml of Sterile Water For Injection I.P. of Sterile Water For Injection I.P. 1.0 ml of reconstituted mixture Neutralizes 1.0 ml of reconstituted mixture Neutralizes 0.60 mg. of Cobra Venom 0.60 mg. of Cobra Venom 0.45 mg. of Krait venom 0.45 mg. of Krait venom 0.60 mg. of Russell’s viper venom 0.60 mg. of Russell’s viper venom 0.45 mg. of Saw scaled viper venom 0.45 mg. of Saw scaled viper venom Nature and concentration Nature and concentration of preservative of preservative : Phenol /Cresol not more than < 0.25% : Phenol /Cresol not more than < 0.25% Storage conditionsStorage conditions :Store in cool dark place, avoid :Store in cool dark place, avoid exposure to sunlight for lyophilized exposure to sunlight for lyophilized
and to and to store between 2-8˚C for liquid store between 2-8˚C for liquid product.product.
HOW MUCH ANTIVENOM IS NEEDED??( TUMBARE & KHADILKAR,JOURNAL OF BOMBAY VETERINARY COLLEGE:2004,12(1&2):9-11)
Snake Average Venom Yield in ml (Liquid)
Average Venom Yield in mg (Lyophilized)
Theoretical dose of antivenom required for treatment(10 ml vial)
Indian Cobra
0.098-1.56 ml(0.435±0.01)
56.4-514.9mg(126.32±4.31)
21
Russell’s Viper
0.177 -1.356 ml
(0.412±0.02)
20 -277.5 mg(75.98±3.66)
13
Indian Common Krait
0.03-0.25 ml(0.101±0.01)
1.25-18.89 mg(8.35±0.64)
2
INDIAN SCENARIO
It is estimated that, India currently produces about 1.8-2.0 million Snake Antivenom vials per year
Almost 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 deaths
Many times the problem is compounded by improper distribution
This results in big demand-supply gap for this life saving drug
INDIAN SCENARIO Many 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
INDIAN SCENARIO India still has very low price as compared to
many other countries It 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. 190 -Rs. 210 per vial in 2011
MY VIEWS Dose 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.
THANK YOU