Efficient Removal of Non-Structural Protein Using Chloroform for Foot-and-Mouth Disease Vaccine Production.

Sun Young Park,Jae-Seok Kim,Sang Hyun Park,Young-Joon Ko,Jung-Min Lee,Jong-Hyeon Park,Choi-Kyu Park,Sim-In Lee,Hyejin Kim,Ah-Young Kim

doi:10.3390/vaccines8030483

Sun Young Park, Jae-Seok Kim + Show 8 more

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https://doi.org/10.3390/vaccines8030483

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Abstract

To differentiate foot-and-mouth disease (FMD)-infected animals from vaccinated livestock, non-structural proteins (NSPs) must be removed during the FMD vaccine manufacturing process. Currently, NSPs cannot be selectively removed from FMD virus (FMDV) culture supernatant. Therefore, polyethylene glycol (PEG) is utilized to partially separate FMDV from NSPs. However, some NSPs remain in the FMD vaccine, which after repeated immunization, may elicit NSP antibodies in some livestock. To address this drawback, chloroform at a concentration of more than 2% (v/v) was found to remove NSP efficiently without damaging the FMDV particles. Contrary to the PEG-treated vaccine that showed positive NSP antibody responses after the third immunization in goats, the chloroform-treated vaccine did not induce NSP antibodies. In addition to this enhanced vaccine purity, this new method using chloroform could maximize antigen recovery and the vaccine production time could be shortened by two days due to omission of the PEG processing phase. To our knowledge, this is the first report to remove NSPs from FMDV culture supernatant by chemical addition. This novel method could revolutionize the conventional processes of FMD vaccine production.

Highlights

Foot-and-mouth disease (FMD) is a highly contagious viral disease of cloven-hoofed animals
Monoclonal and Western blot was performed with anti-FMD virus (FMDV) type O VP1 and anti-FMDV 3B monoclonal antibodies
FMD vaccine purity relates to the level of FMD non-structural proteins (NSPs) in the final product, which should not FMD vaccine purity relates to the level of FMD NSPs in the final product, which should not induce antibodies that would interfere with serological tests used for serological surveillance of virus induce antibodies that would interfere with serological tests used for serological surveillance of virus circulation in vaccinated populations [5]

Summary

Introduction

Foot-and-mouth disease (FMD) is a highly contagious viral disease of cloven-hoofed animals. It is the most serious livestock disease due to the economic consequences of its outbreak. The causative agent, FMD virus (FMDV), belongs to the Aphthovirus genus of the family Picornaviridae. The virus exists in the form of seven different serotypes: O, A, Asia 1, C, and South African Territories (SAT) 1, SAT 2, and SAT 3 [1]. A single-stranded positive RNA virus genome encodes four structural proteins (SPs; VP1, VP2, VP3, and VP4) and several non-structural proteins (NSPs; L, 2A, 2B, 2C, 3A, 3B, 3C, and 3D) [2]. The FMDV rapidly replicates and spreads within the infected animal, among in-contact susceptible animals, and through aerosol [3]

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