28] Methods for binding cells to plastic: Application to solid phase immunoassays for cell-surface antigens

Abstract

This chapter describes the methods for binding cells to plastic and their application to solid phase immunoassays for cell-surface antigens. The developments in cell fusion techniques allow the production of monoclonal antibodies specific for cell-surface antigens and highlight the need for rapid and reproducible large-scale screening test systems to detect such antibodies in cell culture supernatants. Assays, in which the binding of antibody is measured, have an advantage over the test systems that rely on the effector function of bound antibody. The binding of antibody to the cells attached to the plates were compared with the binding obtained with cells in suspension. Cells in suspension were either fresh or fixed by glutaraldehyde in suspension and the attachment of the cells was done by glutaraldehyde alone. The binding of anti-DBA/2 alloantisera to DBA/2 and C57B 1/6 thymocytes was investigated by development with [125I]RaMIg. The binding of an isolated and radioiodinated monoclonal antibody directed against H-2Kb to the cells attached by different means was also compared.