Monoclonal antibodies specific for globin chains

Abstract

Six monoclonal antibodies specific for human globin chains are described. They are produced by stable clones obtained by raising hybridomas using cells of mice immunized with either adult or fetal hemoglobin. Characterization of the antibodies included testing against tetrameric human and other animal hemoglobins, isolated hemoglobin chains, and when indicated, cyanogen bromide fragments. Monoclonals 16-2 and 37-8 are beta-chain specific. Antibody 31-2 recognizes an antigenic determinant common to the alpha and beta subunits. Monoclonal 30-3 recognizes determinants best expressed in the alpha 2 beta 2 tetramer. Antibody 45-1 recognizes a determinant common to beta and gamma subunits, while antibody 51-7 is gamma-chain specific. None of the monoclonal antibodies recognizes mouse hemoglobin, and they display significant differences in binding to hemoglobins of various species. The species-specific reactions and the knowledge of the primary structures of globins allowed deductions about the antigenic sites recognized by two of the monoclonals (16-2 and 45-1). These antihemoglobin monoclonal antibodies will provide useful probes for studying hemoglobin expression in vivo and in vitro.