Development of a 2-Plex Luminex-Based Competitive Immunoassay to Quantify Neutralizing Antibodies Induced by Virus-Like Particles for Human Papillomavirus 16 and 18

Pin Lv,Hongfang Wang,Hongmei Zhu,George Wang

doi:10.1155/2011/272806

Pin Lv, Hongfang Wang + Show 2 more

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https://doi.org/10.1155/2011/272806

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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.

Highlights

Cervical cancer is the second most frequent cancer in women in the world, comprising about 10% of all human cancers [1]
Different ratios of HPV16 L1 virus-like particles (VLPs) or Human papillomavirus (HPV)-18 L1 VLP and #38 or #53 microspheres were used for coupling, respectively
The VLP coupled microspheres were reacted with the same amount of related neutralizing monoclonal antibodies (mAbs)

Summary

Introduction

Cervical cancer is the second most frequent cancer in women in the world, comprising about 10% of all human cancers [1]. In China, a total of 500 million women are at risk of cervical cancer [3]. After more than 30 years of research into the infectious nature of cervical cancer, it is well established that human papillomaviruses (HPVs) are the primary cause for cervical cancer [4]. Most cervical cancer cases are linked to genital infection with HPVs. Up till over 130 different types of HPV have been found, where HPV-16 and HPV18 are of high risk, contribute to over 70% of all cervical cancer cases [5]. Prevention of HPV-16 and HPV18’s infection among women becomes an important issue for scientists and epidemiologists to lower the incidence of cervical cancer

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