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Updated: Sep 20, 2026
Generation of Discriminative Human Monoclonal Antibodies from Rare Antigen-specific B Cells Circulating in Blood
Published on: February 6, 2018
[Preparation and biological activities of monoclonal antibody against p185erbB2]
Mei Han1, Beihai Jiang, Jian Wu
1Department of Biochemistry and Molecular Biology, Peking University School of Oncology, Beijing 100034, China.
Objective:
To produce anti- p185erbB2 monoclonal antibody and investigate its biological activities on cancer cells that overexpress p185erbB2.
Methods:
BALB/c mice were immunized with NIH3T3-erbB2 cell to prepare the anti- p185erbB2 monoclonal antibody. The specificity of the monoclonal antibody 1H3 was identified with ELISA, Western blot, immunoprecipitation and immunohistochemistry assays. The effects of monoclonal antibody 1H3 on the proliferation of cell lines that overexpressed p185erbB2 were measured by MTT assay.
Results:
One strain of monoclonal antibody1H3 was obtained. In ELISA assay, 1H3 specifically reacted with cells overexpressing p185erbB2, but had no reaction with cells not overexpressing p185erbB2. The specificity of the monoclonal antibody 1H3 was verified by immunoprecipitation, Western blot and immunohistochemistry techniques. MTT assay was used to confirm that 1H3 could inhibit the growth of cells that overexpressed p185erbB2 to different extent.
Conclusion:
The monoclonal antibody 1H3 obtained can specifically bind to p185erbB2 and significantly inhibit proliferation of cells overexpressing p185erbB2.
Insights
A novel monoclonal antibody, 1H3, specifically targets and inhibits the proliferation of cancer cells overexpressing p185erbB2. This antibody shows significant potential for cancer therapy by targeting the p185erbB2 protein.
Area of Science:
- Immunology
- Oncology
- Molecular Biology
Background:
- The p185erbB2 oncoprotein is a key target in several human cancers.
- Developing targeted therapies against p185erbB2 is crucial for effective cancer treatment.
Purpose of the Study:
- To generate a monoclonal antibody targeting p185erbB2.
- To evaluate the biological activity of this antibody on cancer cells overexpressing p185erbB2.
Main Methods:
- Immunization of BALB/c mice with NIH3T3-erbB2 cells to produce monoclonal antibodies.
- Specificity validation using ELISA, Western blot, immunoprecipitation, and immunohistochemistry.
- Assessment of anti-proliferative effects via MTT assay on relevant cell lines.
Main Results:
- Successful production of the monoclonal antibody 1H3.
- 1H3 demonstrated high specificity for p185erbB2-overexpressing cells across multiple assays.
- MTT assays confirmed that 1H3 significantly inhibited the proliferation of these cancer cells.
Conclusions:
- Monoclonal antibody 1H3 specifically binds to p185erbB2.
- 1H3 effectively inhibits the proliferation of cancer cells with high p185erbB2 expression.
