title: Variation in IgG1 heavy chain allotype does not contribute to differences in biological activity of two human anti-Rhesus (D) monoclonal antibodies.
abstract: BACKGROUND
  Pooled human anti-Rhesus D antiserum is currently administered for the prevention of RhD alloimmunization. Increased demand, and decreased supply, of donated pooled antiserum has led to the investigation of the suitability of human monoclonal anti-RhD antibodies for use in its place. However, it is unclear which biological properties of monoclonal antibodies are important for function in RhD-positive foetal red cell clearance and the prevention of alloimmunization. Various antibodies behave differently in a number of in vitro assays of biological function.
  
  
  OBJECTIVE
  To compare the function and structure of two human anti-RhD IgG1 monoclonal antibodies which differ in their ability to promote red cell lysis in vitro. In particular to examine whether the functional differences correlate to differences in the IgG1 heavy chain constant region (allotype).
  
  
  METHODS
  We report here the cloning, characterization and re-expression in stable myeloma cell transformants of cDNAs coding for two such antibodies, secreted by the heterohybridoma cell lines ESD-1 (THERAD 03) and LHM 70/45.3 (THERAD 06). The cDNAs were then recombined to exchange portions of the Fc encoding regions and the recombinant antibodies were assayed in vitro to determine RhD-positive red cell-dependent activity.
  
  
  RESULTS
  Recombinant THERAD 03 and 06 antibodies behaved identically to the parent antibodies. The 'inactive' THERAD 06 did not have biological activity reconstituted by exchange with the THERAD 03 Fc regions, nor was THERAD 03 activity abolished by the reciprocal Fc region exchange.
  
  
  CONCLUSIONS
  Human monoclonal anti-RhD antibodies can be cloned and re-expressed in stable cell lines, and exhibit identical properties to the parent antibodies. Differences in biological activity cannot be attributed to differences in IgG1 heavy chain allotype.
authors.affiliation: National Science Laboratory, Scottish National Blood Transfusion Service, Scotland, UK. tpaterso@hgmp.mrc.ac.uk
authors.forename: T
authors.initials: T
authors.lastname: Paterson
authors.forename: J
authors.initials: J
authors.lastname: Innes
authors.forename: L
authors.initials: L
authors.lastname: McMillan
authors.forename: I
authors.initials: I
authors.lastname: Downing
authors.forename: M C
authors.initials: MC
authors.lastname: Carter
mesh_terms: D000911:Antibodies, Monoclonal / Q000032:analysis / Q000235:genetics / Q000276:immunology*; D001483:Base Sequence; D002460:Cell Line / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000032:analysis; D006801:Humans; D007126:Immunoglobulin Allotypes / Q000276:immunology*; D007074:Immunoglobulin G / Q000276:immunology*; D007143:Immunoglobulin Heavy Chains / Q000235:genetics / Q000276:immunology*; D007132:Immunoglobulin Isotypes / Q000276:immunology*; D007147:Immunoglobulin Light Chains / Q000235:genetics / Q000276:immunology; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology; D012204:Rh-Hr Blood-Group System / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D007126:Immunoglobulin Allotypes; D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D007132:Immunoglobulin Isotypes; D007147:Immunoglobulin Light Chains; D011994:Recombinant Proteins; D012204:Rh-Hr Blood-Group System