使用MUC1串联重复序列的重叠20肽进行“表位指纹分析”。

'Epitope fingerprinting' using overlapping 20-mer peptides of the MUC1 tandem repeat sequence.

作者信息

Schol D J, Meulenbroek M F, Snijdewint F G, von Mensdorff-Pouilly S, Verstraeten R A, Murakami F, Kenemans P, Hilgers J

机构信息

Department of Obstetrics and Gynecology, University Hospital Vrije Universiteit, Amsterdam, The Netherlands.

出版信息

Tumour Biol. 1998;19 Suppl 1:35-45. doi: 10.1159/000056503.

DOI:10.1159/000056503

PMID:9422087

Abstract

The ISOBM TD-4 Workshop antibodies 122-177 were tested for reactivity with 20 overlapping MUC1 tandem repeat 20-mer peptides by an ELISA, in order to determine the complete amino acid sequences of the epitopes. Of the 56 antibodies studied, 30 showed specific binding and thus the epitopes were characterized. The epitopes appear to be 'broader' when compared to those deduced from studies using smaller peptides. Interassay variation is remarkably small, allowing for precise grouping of clusters with very similar epitope patterns. Five groups of antibodies show remarkable similarity: BC3 and VU-4-H5; BC4W154, C595 and Mc5; MF06 and B27.29; VU-11-D1 and VU-11-E2; Ma552, VU-3-C6, 7540MR and BC4E549. We have used the term 'epitope fingerprinting' to refer to the 'fine structure' of the epitope with all its essential and flanking amino acids. We believe this method is more precise than the usual epitope mapping with short peptides.

摘要

通过酶联免疫吸附测定（ELISA），检测了国际生物标记物标准化委员会（ISOBM）TD - 4研讨会的抗体122 - 177与20个重叠的MUC1串联重复20聚体肽的反应性，以确定表位的完整氨基酸序列。在所研究的56种抗体中，30种显示出特异性结合，从而对表位进行了表征。与使用较小肽段的研究所推导的表位相比，这些表位似乎“更宽泛”。测定间的变异非常小，使得具有非常相似表位模式的簇能够精确分组。五组抗体显示出显著的相似性：BC3和VU - 4 - H5；BC4W154、C595和Mc5；MF06和B27.29；VU - 11 - D1和VU - 11 - E2；Ma552、VU - 3 - C6、7540MR和BC4E549。我们使用“表位指纹图谱”一词来指代表位的“精细结构”及其所有必需和侧翼氨基酸。我们认为这种方法比通常用短肽进行的表位作图更精确。