title: Production and characterization of anti-kell monoclonal antibodies using transfected cells as the immunogen.
abstract: Monoclonal antibodies (Mabs) to blood group antigens are valuable as diagnostic reagents for typing red blood cells (RBCs) in the clinical setting, and for structure-function studies of proteins. Here, we report a powerful system that enabled us to produce Mabs to blood group antigens. A murine erythroleukaemia (MEL) cell line expressing Kell protein, a transmembrane glycoprotein that carries a number of clinically relevant antigens, was used as a novel immunogen. Mabs with different specificities to the Kell protein were produced from a single mouse fusion: an anti-Jsb (MIMA-8), and two antibodies (MIMA-9 and MIMA-10) with novel specificities, that reacted with RBCs with the common Kell phenotype but not with RBCs with K+k- or Kp(a+b-) or K0 phenotypes. The non-reactivity with both K+k- or Kp(a+b-) RBCs implied that the epitope was influenced by the molecular changes associated with an absence of the k or Kpb antigens. MIMA-8 is the first example of a Mab anti-Jsb and was used in the clinical laboratory for screening donor RBCs for Js(b-) blood and for typing RBCs from patients even when the RBCs were coated with anti-IgG as is the case in autoimmune haemolytic anaemia. Heavy and light chain variable regions of MIMA-8 were cloned and the sequence is given. This study illustrates the potential of this novel immunization approach for making monoclonal antibodies to blood group antigens.
authors.affiliation: Marion Bessin Liver Research Center, Albert Einstein College of Medicine, Bronx, New York, N.Y., USA.
authors.forename: T H
authors.initials: TH
authors.lastname: Chu
authors.forename: K
authors.initials: K
authors.lastname: Yazdanbakhsh
authors.forename: R
authors.initials: R
authors.lastname: Oyen
authors.forename: E
authors.initials: E
authors.lastname: Smart
authors.forename: M E
authors.initials: ME
authors.lastname: Reid
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D001483:Base Sequence; D002460:Cell Line; D006825:Hybridomas / Q000276:immunology; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007147:Immunoglobulin Light Chains / Q000235:genetics; D007626:Kell Blood-Group System / Q000276:immunology*; D051379:Mice; D008969:Molecular Sequence Data; D014162:Transfection / Q000276:immunology; D014614:Vaccines, Synthetic / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007626:Kell Blood-Group System; D014614:Vaccines, Synthetic