使用酶联免疫吸附测定法对重叠合成肽文库进行线性B细胞表位作图。

Linear B-cell epitope mapping using enzyme-linked immunosorbent assay for libraries of overlapping synthetic peptides.

作者信息

Heuzenroeder Michael W, Barton Mary D, Vanniasinkam Thiru, Phumoonna Tongted

机构信息

Infectious Diseases Laboratories, Institute of Medical and Veterinary Science, Frome Road, PO Box 14, Rundle Mall, Adelaide, SA 5000, Australia.

出版信息

Methods Mol Biol. 2009;524:137-44. doi: 10.1007/978-1-59745-450-6_10.

DOI:10.1007/978-1-59745-450-6_10

PMID:19377942

Abstract

The aim of this chapter is to provide a strategy for mapping linear antibody epitopes of protein antigens in order to discover candidates for vaccines or diagnostic tests. A set of overlapping peptides was designed and synthesised based upon a known amino acid sequence of the target protein, virulence-associated protein A (VapA) of the bacterium Rhodococcus equi, an important pulmonary pathogen in foals.The peptides were biotinylated and used in an ELISA to screen immune sera from foals. These biotinylated peptides were coated directly onto micro titre plates that had been pre-coated with NeutrAvidin. A linear B-cell epitope was identified by a universal recognition of sera to the synthetic peptides which corresponds to a particular fragment of the VapA protein.

摘要

本章的目的是提供一种绘制蛋白质抗原线性抗体表位的策略，以便发现疫苗或诊断测试的候选物。基于目标蛋白马红球菌（驹类重要的肺部病原体）的毒力相关蛋白A（VapA）的已知氨基酸序列，设计并合成了一组重叠肽。这些肽被生物素化，并用于酶联免疫吸附测定（ELISA）以筛选驹的免疫血清。这些生物素化肽直接包被在预先包被有中性抗生物素蛋白的微量滴定板上。通过血清对合成肽的普遍识别鉴定出一个线性B细胞表位，该合成肽对应于VapA蛋白的一个特定片段。