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由鼠单克隆抗体所界定的人类前列腺组织抗原

Human prostate tissue antigens defined by murine monoclonal antibodies.

作者信息

Starling J J, Sieg S M, Beckett M L, Wirth P R, Wahab Z, Schellhammer P F, Ladaga L E, Poleskic S, Wright G L

出版信息

Cancer Res. 1986 Jan;46(1):367-74.

PMID:2415250
Abstract

BALB/c mice were hyperimmunized against a membrane preparation derived from a pool of transurethral resection specimens which included three benign prostatic hyperplasia and one prostate adenocarcinoma tissue samples. The activated lymphocytes were fused with the NS-1 mouse myeloma cell line, and supernatants from immunogen-reactive hybridomas were screened for antibody binding activity using a solid-phase radioimmunoassay against the Calu-1 human lung adenocarcinoma cell line and several membrane preparations derived from various normal human tissues. Hybridoma cultures secreting antibodies which did not appear cross-reactive were doubly cloned by limiting dilution and screened against a large panel of membrane preparations derived from normal prostate, benign prostatic hyperplasia, and prostate adenocarcinoma tissues as well as samples obtained from a variety of normal human tissues. The monoclonal antibodies were also evaluated against 24 normal, virally transformed, and malignant human cell lines. Two monoclonal antibodies were isolated which demonstrated a restricted binding activity to prostate antigens and were not widely cross-reactive with nonprostate normal tissues or cell lines. These antibodies were designated TURP-27 (IgG3, k) and TURP-73 (IgG2a, k). Both of these monoclonal antibodies were reactive against formalin-fixed, paraffin-embedded tissues in the immunoperoxidase assay and were subsequently tested against a variety of normal, hyperplastic, and malignant human tissues. These studies indicated that TURP-27 may be directed against a new prostate organ-associated marker and that TURP-73 is directed against an antigen expressed on prostate and a limited number of other tissues.

摘要

用来自经尿道前列腺切除术标本池的膜制剂对BALB/c小鼠进行超免疫,该标本池包含三个良性前列腺增生和一个前列腺腺癌组织样本。将活化的淋巴细胞与NS-1小鼠骨髓瘤细胞系融合,使用针对Calu-1人肺腺癌细胞系和几种源自各种正常人体组织的膜制剂的固相放射免疫测定法,筛选免疫原反应性杂交瘤的上清液中的抗体结合活性。通过有限稀释对分泌未出现交叉反应抗体的杂交瘤培养物进行双重克隆,并针对来自正常前列腺、良性前列腺增生、前列腺腺癌组织以及从各种正常人体组织获得的样本的大量膜制剂进行筛选。还针对24种正常、病毒转化和恶性人类细胞系评估了单克隆抗体。分离出两种单克隆抗体,它们对前列腺抗原有受限的结合活性,并且与非前列腺正常组织或细胞系没有广泛的交叉反应。这些抗体被命名为TURP-27(IgG3,κ)和TURP-73(IgG2a,κ)。在免疫过氧化物酶测定中,这两种单克隆抗体均与福尔马林固定、石蜡包埋的组织反应,随后针对各种正常、增生和恶性人类组织进行测试。这些研究表明,TURP-27可能针对一种新的前列腺器官相关标志物,而TURP-73针对前列腺和少数其他组织上表达的一种抗原。

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