title: Development of a 2-plex luminex-based competitive immunoassay to quantify neutralizing antibodies induced by virus-like particles for human papillomavirus 16 and 18.
abstract: Human papillomavirus (HPV) L1 virus-like particles (VLPs) were proven an effective vaccine candidate to prevent against HPV-16 and -18 infections. In order to evaluate the potency of our produced HPV-16 and -18 L1 VLPs-based vaccine candidates, also to quantify neutralizing antibodies induced by them, a 2-plex Luminex-based competitive immunoassay was developed. Unlike the published paper, the no-biotin conjugated neutralizing mAbs spiked normal human serum (NHS) was used for standard curve preparation, while phycoerythrin (PE) was not labeled directly to neutralizing mAbs for signaling. After the coupling optimization of VLPs to microspheres and the neutralizing mAbs biotinylation, the 2-plex standard curve was prepared with good fit and high dynamic range. In addition, no cross-reactivity was also confirmed. The 2-plex Luminex-based immunoassay represents good potential not only for vaccine candidate's evaluation but also for its further clinical use.
authors.affiliation: Analytical Science & Formulation Department, Shanghai Zerun Biotechnology Co., Ltd. and Genor Biopharma Co. Ltd., 1399 Zhangheng Road, Zhangjiang, Shanghai, China.
authors.forename: Pin
authors.initials: P
authors.lastname: Lv
authors.forename: Hongmei
authors.initials: H
authors.lastname: Zhu
authors.forename: Hongfang
authors.initials: H
authors.lastname: Wang
authors.forename: George
authors.initials: G
authors.lastname: Wang
mesh_terms: D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing / Q000032:analysis* / Q000096:biosynthesis / Q000276:immunology; D001426:Bacterial Proteins; D001667:Binding, Competitive; D019921:Biotinylation; D006735:Horseradish Peroxidase; D052162:Human papillomavirus 16 / Q000276:immunology*; D052161:Human papillomavirus 18 / Q000276:immunology*; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D008863:Microspheres; D053918:Papillomavirus Vaccines / Q000276:immunology; D014771:Virion / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D001426:Bacterial Proteins; D053918:Papillomavirus Vaccines; C058865:Strep-avidin conjugated horseradish peroxidase; C492959:human papillomavirus type 18 L1 VLP vaccine; D006735:Horseradish Peroxidase