Abstract
A cell separation technique is described which can provide large numbers (i.e. >5 × 10 7) of human lymphoid cells which are positively and negatively selected for their ability to react with the OKT4 monoclonal antibody. Peripheral blood lymphocytes are depleted of plastic-adherent cells and then exposed to OKT4 antibody. Unbound OKT4 antibody is removed by washing and then this OKT4-exposed cell population is placed on plastic Petri dishes that are coated with affinity-purified goat anti-mouse immunoglobulin. Cells which do not bind the OKT4 antibody (OKT4 − cells) can be obtained by gently washing the nonadherent cells from the plates, while the adherent (OKT4 +) cells can be recovered by vigorous pipetting. This procedure yields functional OKT4 + and OKT4 − cell populations with a purity which approaches that obtained by separation using the fluorescence-activated cell sorter. The principal advantages of this plate separation technique are that: (1) large numbers of functional cell populations can be obtained; (2) very small quantities of monoclonal antibody (e.g. 1:10 4 dilution of ascites) are required; and (3) sterility of the cell preparations can be maintained quite easily.
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