[Preparation and characterization of monoclonal antibody to gamma seminoprotein]
| Autor: | Awato Fujino, Kenjiro Yasuda, Takashi Sogo, Akira Ishibashi, Osamu Hoshiai, Ken Koshiba, Sadakazu Aiso |
|---|---|
| Rok vydání: | 1990 |
| Předmět: |
Male
medicine.drug_class Urology Adenocarcinoma Monoclonal antibody Immunoenzyme Techniques Mice Antigen Prostate Antibody Specificity medicine Animals Humans Mice Inbred BALB C Chemistry Seminal Plasma Proteins Antibodies Monoclonal Prostatic Neoplasms Prostatic Secretory Proteins Proteins medicine.disease Negative stain Molecular biology Transitional cell carcinoma medicine.anatomical_structure Immunohistochemistry Clone (B-cell biology) |
| Zdroj: | Nihon Hinyokika Gakkai zasshi. The japanese journal of urology. 81(1) |
| ISSN: | 0021-5287 |
| Popis: | Gamma seminoprotein (gamma Sm), a glycoprotein isolated from human seminal plasma with a molecular weight of 29,000 and possibly a serine protease, has been demonstrated to be one of the prostate organ-specific antigens. We established a murine monoclonal antibody (MoAb) to gamma-Sm in order to prove the presence and localization of this protein in the prostate. The hybrid clones were obtained by fusing mouse SP2/O-Ag-14 myeloma cells with splenocytes from Balb/c mouse immunized with the major fractions of gamma-Sm. The enzyme-linked immunosorbent assay was done for antibody screening. After cloning twice in soft agarose, the stable clone, termed 43-21-1-1, was finally chosen. This MoAb, IgG1(kappa), recognized gamma-Sm specifically, which was verified by an immunoblotting assay. The specificity of the MoAb was further evaluated by immunohistochemical study by the avidin biotin complex method. Periodate-lysine-paraformaldehyde-fixed surgical specimens, including the prostate associated with fibromuscular hyperplasia, seminal vesicles, bladder, testis and epididymis, were examined. Formaldehyde (10%)-fixed surgical specimens from patients with adenocarcinoma of the prostate and primary transitional cell carcinoma arising from the periurethral prostatic ducts were also examined. Positive reactions of gamma-Sm were recognized only in the cytoplasm of prostatic glandular epithelial cells and along the luminal surface. Fibrous and muscular tissues always given negative staining. Neither nonprostatic tissues nor transitional cell carcinoma of the prostate were stained positively for gamma-Sm. These results show that this MoAb (43-21-1-1) is quite specific to gamma-Sm and may be useful for the immunohistochemical study with prostatic tissue. |
| Databáze: | OpenAIRE |
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