验证基于等离子体的生物传感器在生物样品中快速和深入地表征针对兔出血性和口蹄疫病毒的单克隆抗体。

IF 4.2 3区 生物学 Q1 BIOCHEMICAL RESEARCH METHODS
Chiara Urbinati , Giulia Pezzoni , Patrizia Cavadini , Vittoria Di Giovanni , Lorenzo Capucci , Marco Rusnati
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引用次数: 0

摘要

ELISA和RT-PCR是生物样品中病毒敏感鉴定的标准工具,但它们缺乏精细表征病毒或病毒抗原与单克隆抗体(mab)结合的能力。生物传感技术作为一种强大的单克隆抗体鉴定工具,在病毒学领域正变得越来越重要。表面等离子体共振(SPR)是一种光学生物传感技术,已被用于深入表征具有诊断和治疗价值的单克隆抗体。兔出血性疾病病毒(RHDV)和口蹄疫病毒(FMDV)是兽医面临的首要问题,因此开发针对抗病毒单克隆抗体特征的新方法是具有重要牲畜保健和经济意义的优先事项。考虑到这些前提,在这里,我们通过不同的策略固定RHDV2或其6S亚基制备了一系列SPR生物传感器,然后用于表征抗RHDV2单克隆抗体的结合能力。结果表明,以捕集剂-单抗共价固定在表面捕获的完整RHDV2组成的生物传感器的分析性能最好。为了评估生物传感器的通用性,然后在细胞提取物中使用FMVD采用相同的策略。本文讨论了SPR在动物病毒学诊断和治疗领域中对抗病毒单克隆抗体的快速和弹性精细鉴定中的应用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Validation of plasmonic-based biosensors for rapid and in depth characterization of monoclonal antibodies directed against rabbit haemorrhagic and foot-and-mouth disease viruses in biological samples

Validation of plasmonic-based biosensors for rapid and in depth characterization of monoclonal antibodies directed against rabbit haemorrhagic and foot-and-mouth disease viruses in biological samples
ELISA and RT-PCR represent the standard tools for the sensitive identification of viruses in biological samples, but they lack the capacity to finely characterize the binding of viruses or viral antigens to monoclonal antibodies (MAbs). Biosensing technologies are gaining increasing importance as powerful MAb characterization tools in the field of virology. Surface plasmon resonance (SPR) is an optical biosensing technology already used for the in depth characterization of MAbs of diagnostic and therapeutic value. Rabbit haemorrhagic disease virus (RHDV) and foot-and-mouth disease virus (FMDV) are top veterinary issues for which the development of novel methods aimed at the characterization of antiviral MAbs represents a priority with important livestock healthcare and economic implications. With these premises in mind, here we prepared a series of SPR biosensors by immobilizing RHDV2 or its 6S subunit by different strategies that were then used to characterize the binding capacity of a panel of anti-RHDV2 MAbs. From the comparison of the results obtained, the biosensor composed of intact RHDV2 captured with catcher-MAb covalently immobilized to the surface showed the best analytical performances. To evaluate the versatility of the biosensor, the same strategy was then adopted using FMVD in cell extracts. The results obtained are discussed in view of the exploitation of SPR in the rapid and resilient fine characterization of antiviral MAbs for diagnostic or therapeutic purposes in the field of animal virology.
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来源期刊
Methods
Methods 生物-生化研究方法
CiteScore
9.80
自引率
2.10%
发文量
222
审稿时长
11.3 weeks
期刊介绍: Methods focuses on rapidly developing techniques in the experimental biological and medical sciences. Each topical issue, organized by a guest editor who is an expert in the area covered, consists solely of invited quality articles by specialist authors, many of them reviews. Issues are devoted to specific technical approaches with emphasis on clear detailed descriptions of protocols that allow them to be reproduced easily. The background information provided enables researchers to understand the principles underlying the methods; other helpful sections include comparisons of alternative methods giving the advantages and disadvantages of particular methods, guidance on avoiding potential pitfalls, and suggestions for troubleshooting.
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