Development and application of a method for quantitation of recombinant prefusion F protein of respiratory syncytial virus based on bio-layer interferometry technology
10.13200/j.cnki.cjb.004650
- VernacularTitle:基于生物膜干涉技术的重组呼吸道合胞病毒融合前F蛋白含量检测方法的建立及应用
- Author:
Wenxi HUANG
1
Author Information
1. Guangzhou Patronus Biotechnology Co., Ltd., Luye Life Sciences Group, Guangzhou 510320, Guangdong Province, China
- Publication Type:Journal Article
- Keywords:
Respiratory syncytial virus(RSV);
Bio-layer interferometry(BLI);
Prefusion F protein(Pre-F);
Trimer
- From:
Chinese Journal of Biologicals
2026;39(02):208-216
- CountryChina
- Language:Chinese
-
Abstract:
Objective To develop an analytical method for quantitative detection of prefusion F protein(Pre-F) of respiratory syncytial virus(RSV) based on bio-layer interferometry(BLI) technology, and preliminarily verify and apply the method in order to provide key technical supports for production process development and quality control.Methods RSV Pre-F protein specific antibodies were screened as detection antibody, and Protein G biosensor was used to establish a quantitative assay for RSV Pre-F protein based on BLI technology. The linear range, specificity, accuracy, precision, sensitivity and dilution reliability of the assay were investigated to preliminarily confirm its performance. In addition, the samples from different stages of manufacturing process of recombinant RSV protein vaccine were detected by the BLI assay, and the results were compared with those obtained by enzyme linked immunosorbent assay(ELISA) and high-performance liquid chromatography(HPLC).Results RSV Pre-F trimer protein specific antibody AM14 was selected as the detection antibody,the indirect detection mode of “Protein G sensor-AM14 antibody-Pre-F protein” was constructed, and the BLI assay was established. The linear range of the assay was 1. 25-40 ??g/mL, and only the intact trimer Pre-F protein could be detected,showing high specificity. The recovery rate in the accuracy verification was between 80% and 120%, and the CVs of repeatability and intermediate precision verification were less than 15%. The limit of quantitation was 1. 25 ??g/mL. The assay showed good repeatability and reliability in the samples from all stages of manufacturing process. The assay was suitable for the detection of Pre-F protein in samples at all stages of recombinant RSV protein vaccine manufacturing process, and was comparable to ELISA method while superior to HPLC method.Conclusion The BLI assay for quantitative detection of RSV Pre-F protein has been successfully established. The assay has high specificity, good linearity, accuracy,precision, sensitivity and dilution reliability, and has advantages in simple and rapid operation and tolerance to various complex substrates.
