Related Experiment Video
Updated: Aug 12, 2026
Generation of Recombinant Human IgG Monoclonal Antibodies from Immortalized Sorted B Cells
Published on: June 5, 2015
[Characterization and validation of a human anti-C monoclonal antibody]
Insights
A new monoclonal IgM antibody specific for the Rh C antigen was developed. This potent reagent has been successfully used for phenotyping 30,000 patients and pregnant women.
Area of Science:
- Immunology
- Serology
- Blood banking
Background:
- Development of specific reagents for blood group antigen detection is crucial for transfusion medicine.
- Monoclonal antibodies offer high specificity and reproducibility in serological testing.
Observation:
- Peripheral lymphocytes from a woman immunized against Rh C and Ce antigens were used to generate a hybridoma.
- The resulting hybridoma produced a monoclonal IgM antibody with specificity for the C determinant of the Rh system.
Findings:
- Serological characterization confirmed the antibody's specificity for the C antigen, showing strong reactions with C-positive red blood cells via saline and enzymatic methods.
- Validation using automated hemagglutination tests against 2,500 patient samples showed no discrepancies, confirming its reliability.
Implications:
- This monoclonal IgM anti-C antibody serves as a potent and reliable reagent for Rh C antigen phenotyping.
- Successful application in phenotyping approximately 30,000 individuals demonstrates its clinical utility in blood banking and transfusion practices.
Abstract:
Peripheral lymphocytes were obtained from an immunized woman against C and Ce antigens. Punction was realized 14 days after childbirth. After hetero-hybridization, a unique cell line continued secreting a monoclonal IgM antibody. Serological characterization of this antibody was determined by direct agglutination tests against 150 native and enzyme treated red blood cells including some rare phenotypes. This antibody was specific of C determinant of the Rh system. It showed strong reactions by saline and enzymatic technics, against C positive cells from Ce positive and Ce negative patients. The validation was performed by a manual direct agglutination test in saline (tube) and by an automated-hemagglutination test (in microplate) against 2,500 patients samples. No discrepancy was observed. This monoclonal IgM anti-C could be used as a potent reagent and since one year, about 30,000 patients and pregnant women have been phenotyped in our laboratory successfully.
Related Concept Videos
01:31Hybridoma Technology
Hybridoma Selection
Commonly used fusion techniques — electroporation, polyethylene glycol...
01:25Antibody Structure and Classes
The basic structure of an antibody consists of four protein chains: two identical heavy chains and two identical light chains. These chains are held together by disulfide bonds and other non-covalent interactions, forming a Y-shaped structure.