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Integrity of GH-loop of foot-and-mouth disease virus during virus inactivation: detection by epitope specific
P K Patil1, V Suryanarayana, Pradeep Bist
1Indian Veterinary Research Institute, Hebbal Campus, 560024, Bangalore, India.
Abstract:
Vaccine against foot-and-mouth disease (FMD) is prepared after inactivating the virus produced in cell culture. Inactivation of the FMD virus (FMDV) was earlier done by formaline. However, several vaccine outbreaks, which occurred in Europe revealed that the formaline treatment is not highly effective for virus inactivation. Subsequently, binary ethyleneimine (BEI) was identified as an effective inactivation reagent for FMDV. However, these chemical reagents are likely to have effect on whole virus particle whose integrity is essential for vaccine potency. Therefore, a need is felt to develop non-chemical methods. We have studied induction of endonucleolytic activity as an alternative method for virus inactivation. This method of inactivation was compared with the chemical methods, and found to be highly effective for virus inactivation. The effects of endonucleolytic activity on the integrity of virus capsid was studied using antibodies raised against recombinant proteins, which elicited antibodies against major epitopes present on the surface of the virus. Further, the effect of the agents on the integrity of the virus capsid was studied by using antigen capture PCR (Ag-RT/PCR) which detects the whole virus. The studies showed that inactivation of the virus by induction of endonucleolytic activity is more effective besides maintaining virus integrity. The effect of various inactivating agents on four serotypes of FMDV has also been studied and found to have varying effects, depending on serotype.
Insights
Foot-and-mouth disease virus (FMDV) inactivation for vaccines is improving. Endonucleolytic activity offers a more effective, non-chemical method than traditional formalin or binary ethyleneimine (BEI), preserving virus integrity for enhanced vaccine potency.
Area of Science:
- Veterinary Virology
- Vaccine Development
- Molecular Biology
Background:
- Traditional Foot-and-mouth disease virus (FMDV) inactivation methods using formalin or binary ethyleneimine (BEI) have limitations.
- Concerns exist regarding the impact of chemical inactivating agents on FMDV capsid integrity, potentially affecting vaccine efficacy.
- Previous FMDV inactivation strategies have faced challenges, necessitating the exploration of alternative, non-chemical approaches.
Purpose of the Study:
- To investigate the efficacy of inducing endonucleolytic activity as a novel method for FMDV inactivation.
- To compare the effectiveness of endonucleolytic inactivation with conventional chemical methods (formalin, BEI).
- To assess the impact of endonucleolytic inactivation on the integrity of the FMDV capsid and its implications for vaccine potency.
Main Methods:
- Induction of endonucleolytic activity was studied as a non-chemical virus inactivation strategy.
- FMDV inactivation was compared between endonucleolytic and chemical (formalin, BEI) methods.
- Virus capsid integrity was evaluated using antibodies against recombinant viral proteins and antigen capture PCR (Ag-RT/PCR).
Main Results:
- Induction of endonucleolytic activity proved highly effective for FMDV inactivation.
- This method demonstrated superior effectiveness compared to traditional chemical inactivation techniques.
- Endonucleolytic activity maintained the integrity of the virus capsid, essential for vaccine potency.
- Studies on four FMDV serotypes revealed varying inactivation effects depending on the specific serotype.
Conclusions:
- Induction of endonucleolytic activity represents a promising, non-chemical alternative for FMDV inactivation in vaccine production.
- This method enhances inactivation efficacy while preserving critical virus capsid integrity.
- Further research into serotype-specific effects is warranted for optimized FMDV vaccine development.
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