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Isolation and characterization of human B cell alloantigens
Journal of Immunology (Baltimore, Md. : 1950)
|November 1, 1976
Summary
Researchers identified human B lymphocyte alloantigens from lymphoma cells. These antigens, when purified, revealed distinct molecular weights and potential subunit structures, aiding in understanding B cell surface markers.
Area of Science:
- Immunology
- Cell Biology
- Biochemistry
Background:
- Human B lymphocyte alloantigens are crucial cell surface markers.
- Understanding their structure aids in diagnosing and treating B cell malignancies like lymphoma.
Purpose of the Study:
- To solubilize and characterize human B lymphocyte alloantigens from lymphoma cells.
- To determine the molecular weight and subunit composition of these antigens.
Main Methods:
- Solubilization of B lymphocyte alloantigens using detergent or papain.
- Purification via chromatography (QAE-A50 Sephadex, Concanavalin A Sepharose 4B) and preparative SDS-PAGE.
- Immunoprecipitation of cell line extracts to identify polypeptide subunits.
Main Results:
- Papain-extracted B antigen had a molecular weight of 58,000 Da, while detergent-extracted was 65,000 Da.
- Antigenic activity was consistently detected by both rabbit and human antisera.
- Immunoprecipitation revealed two B cell polypeptides (27,000 and 35,000 Da) potentially forming the native alloantigen.
Conclusions:
- The study successfully isolated and began characterizing human B lymphocyte alloantigens.
- Evidence suggests the 65,000 Da alloantigen is composed of non-covalently linked subunits.
- These findings provide insights into the molecular basis of B cell alloantigen recognition.