Hosoda K, Okada M, Hirata Y, Saito T, Arisato N, Ishigaki S, Niitsu Y
Teijin Institute for Biomedical Research, Tokyo, Japan.
Hybridoma. 1990 Apr;9(2):133-42. doi: 10.1089/hyb.1990.9.133.
Four independent antibodies (6A, 5F, 2H and 2F) to Glutathione S-transferase pi (GST-pi) were selected to characterize their epitopes. Amino acid analysis of 5.6 K and 7.4 K tryptic peptides appeared to suggest that the epitope recognized by antibodies 2H and 2F is located in the N-terminal 44 peptides of GST-pi, and that of 6A and 5F is located in the C-terminal 69 peptides. Reactivities of antibodies 6A and 5F with two synthetic peptides indicated that 6A recognized an epitope in the C-terminal hydrophilic fragment 176Leu-209Gln, and could be distinguished from 5F which recognized an epitope in the 141Thr-176Leu hydrophobic fragment. The differential immuno-reactivity of antibodies 6A and 5F with GST-pi itself, was characterized by the particularly high reactivity of 6A and almost no reactivity of 5F with the natural conformation of GST-pi in solution. This may be explained by differences in the hydropathic natures of their epitopes. The 6A antibody was useful for immunodetection of GST-pi in circulation, while 5F was found to be most suitable for histochemical staining of tumor tissue.
选择了四种针对谷胱甘肽S-转移酶pi（GST-pi）的独立抗体（6A、5F、2H和2F）来表征它们的表位。对5.6K和7.4K胰蛋白酶肽段的氨基酸分析似乎表明，抗体2H和2F识别的表位位于GST-pi的N端44个肽段中，而6A和5F的表位位于C端69个肽段中。抗体6A和5F与两种合成肽的反应性表明，6A识别C端亲水性片段176Leu-209Gln中的一个表位，并且可以与识别141Thr-176Leu疏水性片段中一个表位的5F区分开来。抗体6A和5F与GST-pi本身的免疫反应性差异表现为，6A具有特别高的反应性，而5F对溶液中GST-pi的天然构象几乎没有反应性。这可能是由于它们表位的亲水性性质不同所致。6A抗体可用于循环中GST-pi的免疫检测，而发现5F最适合肿瘤组织的组织化学染色。
