Koyama K
Department of Obstetrics and Gynecology, Hyogo Medical College.
Nihon Sanka Fujinka Gakkai Zasshi. 1988 Aug;40(8):1044-9.
Since the first successful application of somatic cell fusion by Kohler and Milstein to generate hybridomas secreting monoclonal antibodies (Mabs), this hybridoma technique has been widely applied for production of Mabs against a wide variety of antigens. In the lecture, general procedures for establishment of hybridomas and characteristic properties of Mabs were briefly explained and followed by presentation of our clinical studies using Mabs produced in our laboratory to hCG, sperm and tumor cells. 1. Monoclonal antibodies to hCG. Three hybridomas (5D4, 6E4, 2F8) were established by fusing mouse myeloma cells (P3U1) with spleen cells from BALB/c mice immunized with partially purified hCG. Mabs were obtained either from hybridoma culture media or from ascites of mice inoculated the hybridoma cells intraperitoneally. Three Mabs obtained were all IgG1 and each had a unique binding specificity to hCG, hCG-beta, hCG-alpha and LH. They were applied for development of specific, sensitive and easy to perform hCG assays. A reverse passive hemagglutination assay of urinary hCG with a sensitivity of 12.5 IU/l and a sandwich-enzyme immunoassay of serum hCG with a sensitivity of 0.1 mIU/ml were presented. 2. Monoclonal antibodies to human spermatozoa. For elucidation of mechanisms for induction of auto- or iso-sperm immunization and infertility by antisperm antibodies, antigen analysis of seminal components was essential. For this purpose, several Mabs with strong sperm immobilizing (SI) and agglutinating (SA) activities were generated in mice, rats and humans and the corresponding antigens were analysed.(ABSTRACT TRUNCATED AT 250 WORDS)
自科勒和米尔斯坦首次成功应用体细胞融合技术产生分泌单克隆抗体(Mab)的杂交瘤以来,这种杂交瘤技术已被广泛应用于生产针对多种抗原的单克隆抗体。在讲座中,简要解释了建立杂交瘤的一般程序和单克隆抗体的特性,随后介绍了我们使用实验室生产的针对人绒毛膜促性腺激素(hCG)、精子和肿瘤细胞的单克隆抗体进行的临床研究。1. 抗hCG单克隆抗体。通过将小鼠骨髓瘤细胞(P3U1)与用部分纯化的hCG免疫的BALB/c小鼠的脾细胞融合,建立了三种杂交瘤(5D4、6E4、2F8)。单克隆抗体可从杂交瘤培养基中获得,也可从小鼠腹腔接种杂交瘤细胞后产生的腹水中获得。获得的三种单克隆抗体均为IgG1,每种对hCG、hCG-β、hCG-α和促黄体生成素(LH)都有独特的结合特异性。它们被用于开发特异性强、灵敏度高且易于操作的hCG检测方法。展示了灵敏度为12.5 IU/l的尿hCG反向被动血凝试验和灵敏度为0.1 mIU/ml的血清hCG夹心酶免疫试验。2. 抗人精子单克隆抗体。为了阐明抗精子抗体诱导自身或同种精子免疫及不育的机制,精液成分的抗原分析至关重要。为此,在小鼠、大鼠和人类中产生了几种具有强精子固定(SI)和凝集(SA)活性的单克隆抗体,并对相应抗原进行了分析。(摘要截短于250字)