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Updated: Jul 15, 2025

A High Throughput MHC II Binding Assay for Quantitative Analysis of Peptide Epitopes

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Published on: March 25, 2014

扩大MAPPs分析以适应MHC-II泛受体,以提高潜在T细胞表皮质的可预测性

Katharina Hartman¹, Guido Steiner¹, Michel Siegel¹

¹Roche Pharma Research and Early Development, Roche Innovation Center Basel, Grenzacherstrasse 124, 4070 Basel, Switzerland.

|September 28, 2023

概括

这项研究优化了主要基因相容性复合体 (MHC) II相关蛋白质组学 (MAPPs) 试验,用于识别T细胞表位. 特定抗体的组合增强了对人类白细胞抗原 (HLA) II呈现的的检测,改善了免疫性风险评估.

科学领域:

免疫学免疫学免疫学
蛋白质组学是指蛋白质组学.

背景情况:

人类白细胞抗原 (HLA) II呈现对于T细胞免疫反应和生物治疗免疫性至关重要.
目前的主要组织相容性复合体 (MHC) II相关蛋白质组学 (MAPPs) 测定主要使用HLA-DR,限制了从其他HLA II等位基因如HLA-DP和HLA-DQ中识别T细胞表位.
缺乏具有良好特征的免疫沉降抗体,阻碍了在MAPP测定中进行全面的HLA II分析.

研究的目的:

评估商业上可用的MHCII类 (MHC-II) pan,HLA-DP和HLA-DQ在MAPPs测定中的抗体.
为了描述已识别的的结合特异性.
在免疫性风险评估中开发一个优化的MAPP策略,以改进T细胞表皮质检测.

主要方法:

在MAPP测定中测试了商业上可用的抗体克隆 (MHC-II面的CR3/43,WR18,Tü39;HLA-DP的B7/21;HLA-DQ的SPV-L3,1a3)
使用液体染色学-双重质谱法 (LC-MS/MS) 进行鉴定.
基于它们对不同HLA II等位基的结合特异性来表征已识别的.

主要成果:

在基于抗体克隆性的HLA II受体沉试剂性能中发现了显著的变异性,即使在具有类似报告特异性的试剂中也是如此.
证明MHC-II部分抗体不能始终捕获HLA II呈现的的全部光谱.
发现没有单个抗体克隆能够恢复完整的HLA II曲目.

结论:

一个单独的抗体克隆不足以全面恢复HLA II谱.
建议使用L243,WR18和SPV-L3抗体克隆在单个免疫沉阶段的综合策略,以加强和强大的化合物特异性的检测.
优化的MAPPs策略显著提高了T细胞表位的可预测性和鉴定性,从而提高了免疫性风险评估.

关键词:

在 HLA II 受体上.在MAPP测试中,测试方法是:在NetMHCIIpanII上,我们可以在T细胞表观上,T细胞的表观.对抗药物抗体的抗体.免疫性免疫性免疫性免疫类药物免疫类药物在分析中,分析.质谱测量质谱测量质谱测量质谱测量质量测量质谱测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量质量测量治疗性抗体治疗性抗体.

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