Adaptation of an ELISA assay for detection of IgG2a responses against therapeutic monoclonal antibodies in a mouse immunization model.

IF 3.1 4区 医学 Q3 TOXICOLOGY
Shraddha S Rane, Rebecca J Dearman, Ian Kimber, Jeremy P Derrick
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引用次数: 0

Abstract

Biotherapeutic monoclonal antibodies (mAb) play important roles in clinical medicine but their potential to elicit immune responses in patients remains a major issue. In a study designed to investigate the effect of aggregation on immunogenic responses, mice were immunized with two monoclonal antibodies (mAb1 and mAb2). Serum levels of total IgG, IgG1, and IgG2a were measured by ELISA. An anti-mouse IgG2a monoclonal detection antibody cross-reacted with mAb2 but not mAb1, leading to high background when the ELISA plate was coated with mAb2. The problem was solved by use of a goat anti-mouse IgG2a polyclonal antibody that demonstrated the required specificity. IgG2a responses were similar for monomer- or aggregate-coated ELISA plates. The results demonstrate the importance of assessment of the specificity of individual reagents when measuring antibody responses against therapeutic antibodies by ELISA.

ELISA检测IgG2a对治疗性单克隆抗体在小鼠免疫模型中的应答的适应性
生物治疗性单克隆抗体(mAb)在临床医学中发挥着重要作用,但其引发患者免疫反应的潜力仍然是一个主要问题。在一项旨在研究聚集对免疫原性反应影响的研究中,用两种单克隆抗体(mAb1和mAb2)免疫小鼠。ELISA法检测血清总IgG、IgG1、IgG2a水平。抗小鼠IgG2a单克隆检测抗体与mAb2交叉反应,但与mAb1不交叉反应,导致ELISA板包被mAb2时本底高。使用山羊抗小鼠IgG2a多克隆抗体解决了这个问题,该抗体显示了所需的特异性。IgG2a反应在单体或聚合体包被的ELISA板上相似。结果表明,在ELISA测定抗体对治疗性抗体的反应时,评估单个试剂特异性的重要性。
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来源期刊
Journal of Immunotoxicology
Journal of Immunotoxicology 医学-毒理学
CiteScore
6.70
自引率
3.00%
发文量
26
审稿时长
1 months
期刊介绍: The Journal of Immunotoxicology is an open access, peer-reviewed journal that provides a needed singular forum for the international community of immunotoxicologists, immunologists, and toxicologists working in academia, government, consulting, and industry to both publish their original research and be made aware of the research findings of their colleagues in a timely manner. Research from many subdisciplines are presented in the journal, including the areas of molecular, developmental, pulmonary, regulatory, nutritional, mechanistic, wildlife, and environmental immunotoxicology, immunology, and toxicology. Original research articles as well as timely comprehensive reviews are published.
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