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采用针对表位的单克隆抗体进行 ELISA 检测，定量检测循环中的 HER2，有望用于监测接受曲妥珠单抗治疗的癌症患者。

ELISA assay employing epitope-specific monoclonal antibodies to quantify circulating HER2 with potential application in monitoring cancer patients undergoing therapy with trastuzumab.

机构信息

Regional Center for Biomarkers, Department of Clinical Pathology and Transfusion Medicine, Azienda ULSS 3 Serenissima, Regional Hospital, Campo SS Giovanni e Paolo 6777, 30122, Venice (VE), Italy.

Division of Immunology and Allergy, Centre Hospitalier Universitaire Vaudois (CHUV), Rue du Bugnon 46, 1011, Lausanne, Switzerland.

出版信息

Sci Rep. 2020 Feb 20;10(1):3016. doi: 10.1038/s41598-020-59630-y.

DOI:10.1038/s41598-020-59630-y
PMID:32080226
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7033231/
Abstract

Circulating HER2 extracellular domain (HER2 ECD) levels were proposed as a surrogate for HER2 tissue expression to monitor breast cancer patients for early relapse or responses to standard or HER2-targeted therapies, such as the monoclonal antibody (mAb) trastuzumab. Currently, available commercial ELISA assays for HER2 ECD rely on antibodies recognizing undisclosed or unknown epitopes. In this work, two ELISA assays employing MGR2 and MGR3 epitope-specific mAbs for HER2 ECD were developed and validated, showing good assay precision and linearity of the dose-response signal within the dynamic range of 0.19-12.50 ng mL and detection limits of 0.76 and 0.75 ng mL for the MGR2 and MGR3 assays, respectively. The developed assay showed a good agreement with two widely used commercial kits for HER2 ECD quantification in serum samples from breast cancer patients. A complete characterization of mAb-HER2 ECD interaction was performed by means of surface plasmon resonance using trastuzumab as control for both epitope mapping and kinetics analysis. The epitopes recognized by the two mAbs showed no overlap with trastuzumab, which was confirmed by trastuzumab interference analysis in serum samples. The method showed to be a practical approach to determine HER2 ECD with a high degree of sensitivity, reliability and recovery in samples containing mAbs-based therapies.

摘要

循环 HER2 细胞外结构域（HER2 ECD）水平被提议作为 HER2 组织表达的替代物，以监测乳腺癌患者的早期复发或对标准或 HER2 靶向治疗的反应，如单克隆抗体（mAb）曲妥珠单抗。目前，用于 HER2 ECD 的商业 ELISA 检测方法依赖于识别未公开或未知表位的抗体。在这项工作中，开发并验证了两种使用 MGR2 和 MGR3 表位特异性 mAb 用于 HER2 ECD 的 ELISA 检测方法，显示出良好的检测精度和剂量反应信号的线性度，在 0.19-12.50ng/mL 的动态范围内，检测限分别为 MGR2 和 MGR3 检测方法的 0.76 和 0.75ng/mL。开发的检测方法与两种广泛用于乳腺癌患者血清样本中 HER2 ECD 定量的商业试剂盒具有良好的一致性。通过使用曲妥珠单抗作为两种表位作图和动力学分析的对照，利用表面等离子体共振对 mAb-HER2 ECD 相互作用进行了全面表征。两种 mAb 识别的表位与曲妥珠单抗没有重叠，这通过曲妥珠单抗在血清样本中的干扰分析得到了证实。该方法在含有基于 mAb 的治疗药物的样本中具有高度的灵敏度、可靠性和回收率，是一种实用的测定 HER2 ECD 的方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e283/7033231/9ba1666fbc73/41598_2020_59630_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e283/7033231/367304808acc/41598_2020_59630_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e283/7033231/384e14d5be21/41598_2020_59630_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e283/7033231/491279a96d9d/41598_2020_59630_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e283/7033231/3c9ac04e0460/41598_2020_59630_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e283/7033231/08f07332cd1c/41598_2020_59630_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e283/7033231/9ba1666fbc73/41598_2020_59630_Fig6_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e283/7033231/367304808acc/41598_2020_59630_Fig1_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e283/7033231/384e14d5be21/41598_2020_59630_Fig2_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e283/7033231/491279a96d9d/41598_2020_59630_Fig3_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e283/7033231/3c9ac04e0460/41598_2020_59630_Fig4_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e283/7033231/08f07332cd1c/41598_2020_59630_Fig5_HTML.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e283/7033231/9ba1666fbc73/41598_2020_59630_Fig6_HTML.jpg

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