Establishment of Sandwich ELISA for Quality Control in Rotavirus Vaccine Production.

Cao Li,Yuanjun Zeng,Guoxing Luo,Shiyin Zhang,Yingbin Wang,Feibo Song,Tingdong Li,Han Yang,Ningshao Xia,Shengxiang Ge

doi:10.3390/vaccines10020243

Abstract

Non-replicating rotavirus vaccines are alternative strategies that may improve the protective efficacy of rotavirus vaccines in low- and middle-income countries. The truncated spike protein VP4 (aa26-476, VP4*)was a candidate antigen for the development of recombinant rotavirus vaccines, with higher immunogenicity and protective efficacy compared to VP8* and VP5* alone. This article describes the development of three genotype-specific sandwich ELISAs for P[4], P[6], and P[8]-VP4*, which are important for quality control in rotavirus vaccine production. Our results showed that the detection systems had good specificity for the different genotype VP4* and were not influenced by the E. coli host proteins. Moreover, the detection systems play an important role in determining whether the target protein was contaminated by VP4* proteins of other genotypes. They can also detect the adsorption rate of the adjuvant to the P[4], P[6], P[8]-VP4* protein during the process development. The three detection systems will play an important role in the quality control and process development of VP4* based rotavirus vaccines and facilitate the development of recombinant rotavirus vaccines.

Highlights

Rotavirus is the most common cause of severe diarrhea in infants and young children under 5 years [1], responsible for up to 200,000 child deaths each year [2]
We proved that the VP4*stimulated higher titers of neutralizing antibodies and conferred higher protective efficacy than VP8* and VP5* alone, and VP4* could be a viable candidate for rotavirus vaccines [12]
We developed sandwichenzyme-linked immunosorbent assays (ELISA) to detect different genotype VP4* antigens, which could be used for process development and quality control of VP4* antigens

Summary

Introduction

Rotavirus is the most common cause of severe diarrhea in infants and young children under 5 years [1], responsible for up to 200,000 child deaths each year [2]. The protective efficacy of these rotavirus vaccines is decreased in low- and middle-income countries (LMICs), where rotavirus-related mortality is higher [4,5]. Studies have focused on non-replicating rotavirus vaccines (NRRV), which can be administered parenterally and may circumvent the reduced efficacy of live oral rotavirus vaccines [6,7]. A series of rotavirus antigens have been shown to be effective in animal models [8–12]. Among these antigens, the spike protein VP4, which mediates the attachment and penetration of rotavirus infection, was most widely explored [13]. We proved that the VP4*stimulated higher titers of neutralizing antibodies and conferred higher protective efficacy than VP8* and VP5* alone, and VP4* could be a viable candidate for rotavirus vaccines [12]

Methods

Results

Conclusion