Abstract
A local virus isolate, O/SKR/JC/2014 (O JC), has been considered as a candidate vaccine strain in the development of a domestic foot-and-mouth disease (FMD) vaccine in Korea. However, producing and preserving a sufficient quantity of intact vaccine antigens from the O JC strain was difficult owing to its distinctive structural instability compared to other candidate vaccine strains. Based on this feature, the O JC strain was adopted as a model virus for the stabilization study to determine the optimal stabilizer composition, which enables long-term storage of the FMD vaccine antigen in both aqueous and frozen phases. In contrast to O JC vaccine antigens stored in routinely used Tris-buffered or phosphate-buffered saline, those stored in Tris-KCl buffer showed extended shelf-life at both 4 °C and −70 °C. Additionally, the combined application of 10% sucrose and 5% lactalbumin hydrolysate could protect O JC 146S particles from massive structural breakdown in an aqueous state for up to one year. The stabilizer composition was also effective for other FMDV strains, including serotypes A and Asia 1. With this stabilizer composition, FMD vaccine antigens could be flexibly preserved during the general production process, pending status under refrigeration and banking under ultrafreezing.
Highlights
Foot-and-mouth disease (FMD) caused by the foot-and-mouth disease (FMD) virus (FMDV), is a highly contagious vesicular disease of cloven-hoofed animals [1]
Each strain of FMD virus (FMDV) was inoculated in BHK21 suspension cells at a multiplicity of infection (MOI) of 0.002 and was incubated at 37 ◦C in a 5% CO2 shaking incubator at 110 rpm
Identification of FMDV O/SKR/JC/2014 (O JC) as a Highly Unstable Model Virus When the four strains of FMDV, including O JC, O SKR/BE/2017 (O BE), A SKR/YC/2017 (A YC), and As 1 Shamir, we4reof 10 suspended in normal Tris-buffered saline (TBS) and subjected to heating at 45 °C for 30 min, all the other strains, except for O JC, showed a gentle decrease in 146S content as their loss after 30 motihnewr satsraleinss,thexacne2p0t%for(FOigJuCr,es1h)o. wHeodwaevgeern, tOleJdCecdriesapsleayined14a6dSrcaosntitcednet carsetahsei,ralso5ss0%after of th30e minintiawl a1s46leSscsotnhatenn2t0w%as(Fliogsut raeft1e)r. 5Hmowineavnerd, OwaJCs cdoimspplaleyteedlyaddisrsaostciicadteedcraefateser,3a0sm50in% of (Figtuhreein1)i.tiTahl u14s6, iSt cwoansteidnet nwtiafsieldostthafttFeMr 5DmVinOaJnCdhwasacshcaormacptelertiestliyc dinisstaobcialiteydcoafmtepra3r0edmin to o(tFhiegruFreM1D).VThstursa,initsw. as identified that FMDV O JC has characteristic instability compared to other FMDV strains
Summary
Foot-and-mouth disease (FMD) caused by the FMD virus (FMDV), is a highly contagious vesicular disease of cloven-hoofed animals [1]. The FMD vaccine is mainly comprised of inactivated virus particles and adjuvants. Known as 146S particle owing to its sedimentation coefficient, is composed of 60 copies of a protomer. Each of which consists of one set of structural proteins (VP1, VP2, VP3, and VP4), assemble to form a pentamer, and 12 copies of the pentamer and the encapsidated viral RNA form a 146S particle [3]. Intact virus particles, followed by empty capsids (75S) are known to confer the most potent protective immunity to vaccinated animals compared to pentamers (12S) that are poor at conferring immune protection [4]; 146S can dissociate into less immunogenic 12S particles by weak acid or mild heat [4,5]. Stabilization of 146S particles is a key technique for FMD vaccine production
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