Identification of prostate specific membrane antigen (PSMA) as the target of monoclonal antibody 107-1A4 by proteinchip®;: Array, surface-enhanced laser desorption/ionization (SELDI) technology
Identification of prostate specific membrane antigen (PSMA) as the target of monoclonal antibody 107-1A4 by proteinchip®;: Array, surface-enhanced laser desorption/ionization (SELDI) technology
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DOI：
10.1002/ijc.1272
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发表时间：
2001-06-15
影响因子：
6.4
通讯作者：
Vessella, RL
中科院分区：
文献类型：
--
作者：
Wang, SY;Diamond, DL;Vessella, RL
最近，我们报道了前列腺癌组织特异性单抗(MAb)107-IA4的产生、表达模式和对人前列腺癌异种移植瘤的初步靶向。在这份报告中，我们用免疫亲和吸附、SDS-PAGE和凝胶内胰酶消化产生的多肽进行了质谱分析，证明了单抗107-IA4的靶抗原是前列腺特异性膜抗原(PSMA)。已用已知特异性的单抗通过Western blotts确认了该抗原的身份。单抗107-IA4对Western blotts无反应。107-IA4的构象表位位于PSMA的胞外区，在竞争研究中，单抗107-IA4与LNCaP细胞的结合被自身抑制，而不被其他几种抗PSMA单抗抑制，这表明该表位可能是独一无二的。这些结果表明，107-IA4与PSMA外部区域的构象表位发生反应，该表位在本研究中的抗PSMA单抗中是独一无二的。此外，本工作还展示了质谱学解释抗体-配体相互作用的能力。(C)2001年Wiley-Liss，Inc.
Recently we described the generation of the prostate tissue-specific monoclonal antibody(MAb) 107-IA4, its expression pattern and preliminary targeting of human prostate cancer xenografts. In this report we demonstrate that the target antigen for MAb 107-IA4 is prostate-specific membrane antigen (PSMA) using immunoaffinity absorption followed by SDS-PAGE and mass spectrometric analysis of peptides produced by in-gel tryptic digestion. The identity of the antigen has been confirmed by Western blots using MAbs of known specificity. MAb 107-IA4 is not reactive on Western blots. The conformational epitope for 107-IA4 is on the extracellular domain of PSMA, In competition studies, the binding of MAb 107-IA4 to LNCaP cells is inhibited by itself but not by any other of several other anti-PSMA MAbs, suggesting that the epitope may be unique. These results suggest that 107-IA4 is reactive to a conformational epitope in the external domain of PSMA that is unique among the panel of anti-PSMA MAbs in this study. Furthermore this work demonstrates the ability of mass spectroscopy to elucidate antibody-ligand interaction. (C) 2001 Wiley-Liss, Inc.