QUALITY
SNAKE
CONTROL
OF
ANTIVENOM
DR.M.V.KHADILKAR
Technical Director,
Premium Serums & Vaccines Pvt.Ltd.,
Narayangaon,Dist.-Pune
Maharashtra
PRODUCTION PROCESS
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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
DEPENDS
QUALITY
ON
AND
1.The control of source materials
2. Manufacturing procedures
SAFETY
BASIC RAW MATERIALS IN
ANTIVENOM PRODUCTION
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Snake Venoms
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Hyperimmune Plasma
OBJECTIVES OF THE PURIFICATION PROCESS
High safety and efficacy
 No
impairment of the neutralizing
activity during purification
 Maintenance of high purity
 Free of microbial contaminants and
endotoxins
WH
O
Wor
ksho
p,
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
 Proper
of immune response
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
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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
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OTHER METHODS
Use of whole IgG molecule
Papain Digested Fab fragments
Chromatography (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)
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ANTIVENOM SAFETY
Depends 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.
REASONS FOR REACTIONS
Contamination of plasma
 Enzymatic digestion & removal of unwanted
proteins
 Formulation
 Sterility
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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
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MONOVALENT / POLYVALENT –Has its own
advantages/disadvantages
LEVEL OF REFINEMENT-Enzyme digestion,Ion exchange
chromatography/Affinity chromatography. Effect on cost????
VENOM
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QUALITY
GEOGRAPHICAL ANTIGENIC VARIATION
SPECIFICITY/PARASPECIFICITY
CLINICAL MANAGEMENT-OUTCOME DEPENDS ON
EARLY START OF TREATMENT
ADEQUATE DOSES
SUPPORT SYSTEMS
SNAKE ANTIVENOM I.P.
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Name of Product
: Snake Antivenom I.P.
(Polyvalent, Lyophilized/liquid, Equine origin
Enzyme Refined Immunoglobulin)
Description of Product :Lyophilized 20.0 ml Vial
Along with 10.0 ml W.F.I. I.P. OR liquid
10 ml vial
Potency
: When Reconstituted With 10.0 ml
of Sterile Water For Injection I.P.
1.0 ml of reconstituted mixture Neutralizes
0.60 mg. of Cobra Venom
0.45 mg. of Krait venom
0.60 mg. of Russell’s viper venom
0.45 mg. of Saw scaled viper venom
Nature and concentration
of preservative
: Phenol /Cresol not more than < 0.25%
Storage conditions
:Store in cool dark place, avoid
exposure to sunlight for lyophilized and to
store between 2-8˚C for liquid 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
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INDIAN SCENARIO
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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
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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
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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