Establishment and verification of double-antibody sandwich ELISA for detection of glycoprotein E antigen content in recombinant zoster virus-like particle vaccine
10.13200/j.cnki.cjb.004688
- VernacularTitle:重组带状疱疹病毒样颗粒疫苗糖蛋白E抗原含量双抗体夹心ELISA检测方法的建立及验证
- Author:
Hang CHEN
1
Author Information
1. Guangzhou Patronus Biotechnology Co., Ltd., Guangzhou 510320, Guangdong Province, China
- Publication Type:Journal Article
- Keywords:
Varicella-zoster virus(VZV);
Virus-like particle(VLP);
Glycoprotein E;
Double-antibody sandwich ELISA;
Antigen content
- From:
Chinese Journal of Biologicals
2026;39(04):486-493
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish and verify a double-antibody sandwich ELISA method for detecting the content of varicellazoster virus(VZV) glycoprotein E(VZV-gE) antigen in recombinant zoster virus-like particle(VLP) vaccine, so as to provide key technical support for the optimization of production process and the establishment of quality standards.MethodsMultiple anti-VZV-gE monoclonal antibodies were screened by epitope binning analysis to select capture antibody and detection antibody, and the working concentrations of which were determined by chessboard titration. A double-antibody sandwich ELISA was established to detect the VZV-gE antigen content in the drug substance and drug product of the recombinant zoster VLP vaccine. The linear range, specificity, dilution linearity, accuracy, precision and limit of quantitation(LOQ) of the method were investigated. In addition, the VZV-gE antigen content in multiple batches of drug substance and drug product of vaccine was detected using the established method to investigate the consistency of VZV-gE antigen content.ResultsA double-antibody sandwich ELISA for detecting VZV-gE antigen content in the recombinant zoster VLP vaccine was successfully established using ab272686 monoclonal antibody as capture antibody and VZV-M545 monoclonal antibody as detection antibody. This method could specifically identify VZV-gE antigen and had stability-indicating characteristics. The curve of VZV-gEM reference standard fitted well with A_(450)in the range of 0. 781-100 ng/mL, with R~2= 1. The dilution linearity ranges for detecting drug substance and lyophilized drug product were 4 100-262 400 and 200-12 800, respectively.The recovery rates of accuracy verification were all within the range of 80%-120%. Both the coefficients of variation(CVs)in intra-and inter-assay precision verification were less than 20%, while the LOQs for detecting VZV-gE antigen in the drug substance and lyophilized drug product were both 2 ng/mL. The detection results of VZV-gE antigen content in four batches of drug substance and lyophilized drug product met the expectations and showed good consistency.ConclusionThe established double-antibody sandwich ELISA has good specificity, accuracy, precision and linearity, which is suitable for the detection of VZV-gE antigen content in the recombinant zoster VLP vaccine.
