Elimination of Daudi lymphoblasts from human bone marrow using avidin-biotin immunoadsorption

Abstract

Biotinylated antibodies and an avidin-Sepharose 6MB column were utilized in a novel approach to deplete selected cell populations from human bone marrow. Fluorescein-labeled Daudi lymphoblasts were mixed with bone marrow mononuclear cells in a model system, and removal of Daudi cells was quantitatively assessed with an inverted fluorescent microscope. Treatment using the biotinylated monoclonal antibody 2H7 reactive with Bp32 antigen (expressed on Daudi cells) followed by passage over avidin-Sepharose produced greater than two logs of Daudi cell removal from bone marrow. An alternative method was tested by treating cells successively with nonbiotinylated monoclonal antibody and biotinylated goat antimouse immunoglobulin followed by passage over avidin-Sepharose. Up to three logs of Daudi cells could be eliminated from bone marrow with quantitative recovery of hematopoietic progenitors. The use of biotinylated goat antimouse immunoglobulin eliminates the need to prepare a biotin conjugate of each individual monoclonal antibody. The clinical application of cellular immunoadsorption using the avidin-biotin system may prove useful in bone marrow transplantation.