Kuwabara I, Maruyama H, Mikawa Y G, Zuberi R I, Liu F T, Maruyama I N
Department of Cell Biology, Scripps Research Institute, La Jolla, CA 92037, USA.
Nat Biotechnol. 1997 Jan;15(1):74-8. doi: 10.1038/nbt0197-74.
A bacteriophage lambda surface expression system, lambda foo, was used for epitope mapping of human galectin-3. We constructed random epitope and peptide libraries and compared their efficiencies in the mapping. The galectin-3 cDNA was randomly digested by DNase 1 to make random epitope libraries. The libraries were screened by affinity selection using a microtiter plate coated with monoclonal antibodies. Direct DNA sequencing of the selected clones defined two distinct epitope sites consisting of nine and 11 amino-acid residues. Affinity selection of random peptide libraries recovered a number of sequences that were similar to each other but distinct from the galectin-3 sequence. These results demonstrate that a single affinity selection of epitope libraries with antibodies is able to define an epitope determinant as small as nine residues long and is more efficient in epitope mapping than random peptide libraries.
一种噬菌体λ表面表达系统，即λfoo，被用于人半乳糖凝集素-3的表位作图。我们构建了随机表位和肽库，并比较了它们在作图中的效率。半乳糖凝集素-3 cDNA用DNase 1随机消化以制备随机表位库。这些库通过使用包被有单克隆抗体的微量滴定板进行亲和选择来筛选。对所选克隆进行直接DNA测序确定了两个不同的表位位点，分别由9个和11个氨基酸残基组成。随机肽库的亲和选择回收了许多彼此相似但与半乳糖凝集素-3序列不同的序列。这些结果表明，用抗体对表位库进行单次亲和选择能够确定小至9个残基长的表位决定簇，并且在表位作图中比随机肽库更有效。
