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Development of HPV58 type-specific antibodies and detection kit
Preparative Biochemistry & Biotechnology ( IF 2.9 ) Pub Date : 2022-04-24 , DOI: 10.1080/10826068.2022.2065686
Yuying Liu 1, 2, 3 , Yao Zhang 1 , Guifeng Zhang 2 , Zengmin Yang 1 , Yan Wang 1 , Shuming Wu 1 , Dan Chen 1 , Haijiang Zhang 1 , Yongjiang Liu 1
Affiliation  

Abstract

Currently, human papillomavirus (HPV) vaccines are in short supply, so the development of HPV vaccines has a broad market prospect. The 3-, 9-, and 15-valent HPV vaccines developed by ourselves all contain HPV58-derived antigen components. It is important to detect HPV 58 during vaccine production. Here, we introduced a development process of HPV58 type-specific antibodies and a detection kit. Briefly, HPV58 L1-Virus Like Particles (VLPs) were used as antigens to immunize mice, followed by extraction of the ascites to prepare hybridoma cells. After culturing, the supernatants containing secreted antibodies were harvested, purified, and screened to obtain monoclonal antibodies (mAbs). In the pool of attained monoclonal antibodies, we selected 2F7 and 2G7 to evaluate their subtypes, specificity, neutralizing activity, serum competition, binding affinity and gene sequencing. Finally, an enzyme-linked immunosorbent assay (ELISA) detection kit was assembled with 2F7 and 2G7 mAbs which possessed high specificity to HPV58 L1-VLPs. The detection kit developed by 2F7 and 2G7 could be adopted to specifically detect HPV58 L1 protein with good linearity and detection range, which could be widely used in clinical testing and quality control in the production of HPV vaccines.

Abbreviations: BSA: Bovine serum albumin; CDRs: Complementarity-determining regions; CV: Coefficient of variation; DTT: Dithiothreitol; ELISA: Enzyme-linked immunosorbent assay; HAT: Hypoxanthine-aminopterin-thymidine; HPV: Human Papillomavirus; IC50: 50% inhibition rate; IC90: 90% inhibition rate; mAbs: Monoclonal antibodies; VLP: Virus-like particle



中文翻译:

HPV58型特异性抗体及检测试剂盒的研制

摘要

目前,人乳头瘤病毒(HPV)疫苗供不应求,开发HPV疫苗具有广阔的市场前景。自主研发的3价、9价、15价HPV疫苗均含有HPV58来源的抗原成分。在疫苗生产过程中检测 HPV 58 很重要。在此,我们介绍了HPV58型特异性抗体和检测试剂盒的开发过程。简言之,以HPV58 L1病毒样颗粒(VLPs)为抗原免疫小鼠,提取腹水制备杂交瘤细胞。培养后,收集含有分泌抗体的上清液,纯化并筛选以获得单克隆抗体(mAb)。在获得的单克隆抗体库中,我们选择了 2F7 和 2G7 来评估它们的亚型、特异性、中和活性、血清竞争、结合亲和力和基因测序。最后,使用对 HPV58 L1-VLPs 具有高特异性的 2F7 和 2G7 mAb 组装酶联免疫吸附测定 (ELISA) 检测试剂盒。2F7和2G7开发的检测试剂盒可特异性检测HPV58 L1蛋白,具有良好的线性和检测范围,可广泛应用于HPV疫苗生产中的临床检测和质量控制。

缩写: BSA:牛血清白蛋白;CDRs:互补决定区;CV:变异系数;DTT:二硫苏糖醇;ELISA:酶联免疫吸附试验;HAT:次黄嘌呤-氨基蝶呤-胸苷;HPV:人乳头瘤病毒;IC 50:50%抑制率;IC 90 : 90% 抑制率; mAbs:单克隆抗体;VLP:病毒样颗粒

更新日期:2022-04-24
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