Ichiyama K, Ishikawa D, Tanaka Y, Kashiwa T, Koyanagi Y, Handa S, Yamashita A, Fukushi M, Yamamoto N, Taki T
Department of Microbiology, Tokyo Medical and Dental University School of Medicine, Japan.
Viral Immunol. 1999;12(1):57-66. doi: 10.1089/vim.1999.12.57.
We generated a rat monoclonal antibody (mAb W#10) with the ability to neutralize human immunodeficiency virus type 1IIIB (HIV-1IIIB) infection. The epitope recognized by mAb W#10 was defined as R-I-Q-R-G-P-G by enzyme-linked immunosorbent assay (ELISA) with the use of synthetic peptides. The filamentous phage clones displaying random 15-amino-acid peptides on the amino terminus of the pIII coat protein reacting with mAb W#10 were identified with affinity and immunological selection procedures. Thirteen out of 16 selected phage clones contained the G-X-G-R-X-F sequence in the coat protein region representing significant homology to a part of conserved G-P-G-R-A-F sequence in the V3 loop of various HIV-1 strains. In addition, the phage clones included the G-X-G sequence in the sequence detected by synthetic peptides as the recognition site. The selected phage clones were stained by mAb W#10 specifically and were able to compete with mAb binding to cells expressing viral antigens.
我们制备了一种具有中和人类免疫缺陷病毒1型IIIB株（HIV-1IIIB）感染能力的大鼠单克隆抗体（单克隆抗体W#10）。通过使用合成肽的酶联免疫吸附测定（ELISA），单克隆抗体W#10识别的表位被定义为R-I-Q-R-G-P-G。通过亲和及免疫选择程序，鉴定出在pIII外壳蛋白氨基末端展示与单克隆抗体W#10反应的随机15氨基酸肽的丝状噬菌体克隆。16个选定的噬菌体克隆中有13个在外壳蛋白区域含有G-X-G-R-X-F序列，该序列与各种HIV-1毒株V3环中保守的G-P-G-R-A-F序列的一部分具有显著同源性。此外，在合成肽检测的序列中，噬菌体克隆包含G-X-G序列作为识别位点。选定的噬菌体克隆被单克隆抗体W#10特异性染色，并且能够与单克隆抗体结合表达病毒抗原的细胞进行竞争。
