Grauer L S, Charlesworth M C, Saedi M S, Finlay J A, Liu R S, Kuus-Reichel K, Young C Y, Tindall D J
Hybritech Incorporated, San Diego, California 92196-9006, USA.
J Androl. 1996 Jul-Aug;17(4):353-9.
Based on studies indicating that human glandular kallikrein (hK2) mRNA is present in the prostate, we prepared a monoclonal antibody to a synthetic peptide corresponding to the 41-56 region of hK2 to try to identify the hK2 protein. Although prostate-specific antigen (PSA) and hK2 share 80% homology, the 41-56 amino acid sequence of hK2 is only 50% homologous with PSA. A monoclonal antibody, HK1A523, was identified that demonstrates high specificity for hK2. In western blot analysis, the antibody has a 1,000-fold greater sensitivity for the detection of hK2 than for PSA. The antibody was used to probe spent media from the prostate carcinoma cell line, LNCaP. An immunoreactive species was N-terminally sequenced and identified as mature hK2. HK1A523 was also utilized to probe prostate tumor cytosols and seminal fluid where putative forms of hK2 were also identified. The hK2 protein therefore is expressed and secreted from prostate carcinoma cells.
基于一些研究表明人腺激肽释放酶（hK2）mRNA存在于前列腺中，我们制备了一种针对与hK2 41 - 56区域相对应的合成肽的单克隆抗体，试图鉴定hK2蛋白。尽管前列腺特异性抗原（PSA）与hK2有80%的同源性，但hK2的41 - 56氨基酸序列与PSA仅有50%的同源性。我们鉴定出一种单克隆抗体HK1A523，它对hK2表现出高特异性。在蛋白质印迹分析中，该抗体检测hK2的灵敏度比检测PSA高1000倍。该抗体用于探测前列腺癌细胞系LNCaP的培养液。对一种免疫反应性物质进行N端测序并鉴定为成熟的hK2。HK1A523还用于探测前列腺肿瘤细胞溶质和精液，在其中也鉴定出了推测的hK2形式。因此，hK2蛋白由前列腺癌细胞表达并分泌。
