Preservation of B-cell-associated surface antigens by chemical fixation.
Usual methods of chemical fixation preclude examination of cells with most monoclonal antibodies due to alteration or destruction of the surface antigen itself. A method of chemical stabilization and preservation of human B-cell-associated surface antigens is described which facilitates retrospective flow cytometric analysis. This method involves pretreatment of the cells with protease enzyme inhibitors, followed by chemical cross-linking of surface proteins with 2% formalin, and finally blockade of non-specific reactive groups with excess glycine. Once prepared, the expression of pertinent cellular antigens is stable on the cell surface for as long as 4 years. Such methodology could conceivably be used for preparation of cells for longitudinal quality control of monoclonal antibodies or archival storage of patient specimens for retrospective flow cytometric analysis.
Caldwell
C W
CW
Department of Pathology, Ellis Fischel Cancer Center, University of Missouri School of Medicine, Columbia 65203.
D000818:Animals; D000911:Antibodies, Monoclonal; D000954:Antigens, Surface / Q000032:analysis / Q000737:chemistry*; D001402:B-Lymphocytes / Q000276:immunology*; D002417:Cattle; D002453:Cell Cycle / Q000276:immunology; D005434:Flow Cytometry / Q000379:methods*; D005455:Fluorescent Antibody Technique; D006801:Humans; D017493:Leukocyte Common Antigens / Q000032:analysis; D015260:Neprilysin / Q000032:analysis; D017464:Receptors, Complement 3d / Q000032:analysis; D016707:Tissue Fixation
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D017464:Receptors, Complement 3d; D017493:Leukocyte Common Antigens; D015260:Neprilysin