Related Experiment Videos
[Preparation and characterization of monoclonal antibody to gamma seminoprotein]
A Fujino1, A Ishibashi, K Koshiba
1Department of Urology, School of Medicine, Kitasato University, Kanagawa, Japan.
Abstract:
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.
Insights
Researchers developed a specific monoclonal antibody (MoAb) to gamma seminoprotein (gamma Sm), an organ-specific prostate antigen. This MoAb accurately detects gamma Sm in prostate tissues, aiding in diagnostic research.
Area of Science:
- Biochemistry
- Immunology
- Urology
Context:
- Gamma seminoprotein (gamma Sm) is a glycoprotein found in seminal plasma.
- It is recognized as a prostate organ-specific antigen.
- Understanding its presence and localization is crucial for prostate research.
Purpose:
- To develop a murine monoclonal antibody (MoAb) specific for gamma seminoprotein (gamma Sm).
- To validate the specificity and utility of the MoAb (43-21-1-1) for immunohistochemical studies.
- To investigate the presence and localization of gamma Sm within prostate tissue and other organs.
Summary:
- A stable monoclonal antibody (clone 43-21-1-1, IgG1(kappa)) was successfully generated against gamma seminoprotein (gamma Sm).
- Specificity was confirmed via immunoblotting and immunohistochemistry using various fixed tissue specimens.
- The MoAb exclusively identified gamma Sm in the cytoplasm and luminal surface of prostatic glandular epithelial cells, showing no reactivity in non-prostatic tissues or transitional cell carcinoma.
Impact:
- This highly specific MoAb provides a valuable tool for immunohistochemical analysis of prostatic tissue.
- It can aid in distinguishing prostate tissue and potentially in the diagnosis of prostate-related conditions.
- The findings contribute to the understanding of gamma Sm as a prostate-specific biomarker.