Summary report on the ISOBM TD-4 Workshop: Analysis of 56 monoclonal antibodies against the MUC1 mucin - San Diego, Calif., November 17-23, 1996
Summary report on the ISOBM TD-4 Workshop: Analysis of 56 monoclonal antibodies against the MUC1 mucin - San Diego, Calif., November 17-23, 1996
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DOI：
10.1159/000056500
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发表时间：
1998-01-01
期刊：
影响因子：
--
通讯作者：
Hilgers, J
中科院分区：
文献类型：
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作者：
Price, MR;Rye, PD;Hilgers, J
16 个研究小组参加了 ISOBM TD-4 研讨会，其中使用多种靶抗原和 MUC1 粘蛋白相关合成肽和糖肽研究了 56 种针对 MUC1 粘蛋白的单克隆抗体的反应性和特异性。大多数抗体 (34/56) 定义的表位位于 MUC1 粘蛋白核心的 20 个氨基酸串联重复序列内。在其余 22 种抗体中，有证据表明 16 种抗体的表位中涉及碳水化合物残基。免疫原的类型与每种抗体的特异性之间没有明显的关系。通过直接结合测定（例如通过 ELISA 或 BiaCore）或通过确定合成配体抑制抗体结合相互作用的能力，分析合成肽和糖肽与每种抗体的反应性。研究小组在鉴定与 MUC1 蛋白核心内的肽表位反应的抗体方面具有良好的一致性。使用 Pepscan 分析对重叠合成肽的抗体反应性进行表位作图测试，研究组之间的结果基本一致。 MUC1 蛋白核心内表位的主要特征是全部或部分存在 PDTRPAP 亲水序列。碳水化合物表位不太容易表征，在这方面最有用的试剂是限定的寡糖，而不是富含特定碳水化合物部分的纯化粘蛋白制剂。很明显，碳水化合物残基通过调节表位可及性或掩蔽决定簇，或通过稳定 MUC1 蛋白核心内肽表位的首选构象，参与许多表位。总体而言，这些研究强调了各组之间的一致性，而不是暴露出不一致之处，这增强了抗粘蛋白单克隆抗体特异性分析结果的可信度。
Sixteen research groups participated in the ISOBM TD-4 Workshop in which the reactivity and specificity of 56 monoclonal antibodies against the MUC1 mucin was investigated using a diverse panel of target antigens and MUC1 mucin-related synthetic peptides and glycopeptides. The majority of antibodies (34/56) defined epitopes located within the 20-amino acid tandem repeat sequence of the MUC1 mucin protein core. Of the remaining 22 antibodies, there was evidence for the involvement of carbohydrate residues in the epitopes for 16 antibodies. There was no obvious relationship between the type of immunogen and the specificity of each antibody. Synthetic peptides and glycopeptides were analyzed for their reactivity with each antibody either by assay of direct binding (e.g. by ELISA or BiaCore) or by determining the capacity of synthetic ligands to inhibit antibody binding interactions. There was good concordance between the research groups in identifying antibodies reactive with peptide epitopes within the MUC1 protein core. Epitope mapping tests were performed using the Pepscan analysis for antibody reactivity against overlapping synthetic peptides, and results were largely consistent between research groups. The dominant feature of epitopes within the MUC1 protein core was the presence, in full or part, of the hydrophilic sequence of PDTRPAP. Carbohydrate epitopes were less easily characterized and the most useful reagents in this respect were defined oligosaccharides, rather than purified mucin preparations enriched in particular carbohydrate moieties. It was evident that carbohydrate residues were involved in many epitopes, by regulating epitope accessibility or masking determinants, or by stabilizing preferred conformations of peptide epitopes within the MUC1 protein core. Overall, the studies highlight concordance between groups rather than exposing inconsistencies which gives added confidence to the results of analyses of the specificity of anti-mucin monoclonal antibodies.