Preservation of B-cell-associated surface antigens by chemical fixation

Authors

  • Charles W. Caldwell M.D., Ph.D.

    Corresponding author
    1. Department of Pathology, Ellis Fischel Cancer Center, University of Missouri School of Medicine, Columbia, Missouri
    • Department of Pathology, Ellis Fischel Cancer Center, University of Missouri School of Medicine, 115 Business Loop 70 West, Columbia, MO 65203
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Abstract

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 crosslinking 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. © 1994 Wiley-Liss, Inc.

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