title: Effect of the presence or absence of J chain on expression of recombinant anti-Kell immunoglobulin M.
abstract: The aim of this study was to determine the effect of expressing a recombinant anti-Kell immunoglobulin (Ig) M from two cell lines, CH0 and NS0, on its ability to function as a diagnostic antibody. As a polymeric immunoglobulin, IgM is able to directly agglutinate red blood cells (RBCs), making it a useful blood grouping reagent. To simplify expression, recombinant human IgM (rIgM) from NS0 (a mouse myeloma line) and CHO (Chinese hamster ovary line) cells was expressed in the absence of human J chain. Whereas NS0 expresses mouse J chain, rIgM expressed from CH0 cells lack J chain. Although the ability to polymerize resides within the tailpiece of IgM heavy chain, J chain can influence the polymeric state. This in turn could affect the ability of rIgM to bind its antigen. The variable region of the heavy chain of an anti-Kell IgG was grafted onto the constant region of human IgM and co-expressed with light chain derived from the same antibody. rIgM was purified from each cell line and the strength of direct agglutination assessed. Both cell lines produced polymeric rIgM that was able to specifically bind the target antigen and to directly agglutinate RBCs to the same degree. The presence or absence of J chain did not affect the ability of the rIgM to bind the Kell antigen or the strength of agglutination. The presence of J chain is not required for the production of a functional rIgM for use as a diagnostic reagent. CHO and NS0 lines are both suitable for production of such a reagent.
authors.affiliation: Bristol Institute for Transfusion Sciences, NHSBT, Southmead Road, Bristol BS10 5ND, UK. jacky.gilmour@nbs.nhs.uk
authors.forename: J E M
authors.initials: JE
authors.lastname: Gilmour
authors.forename: S
authors.initials: S
authors.lastname: Pittman
authors.forename: R
authors.initials: R
authors.lastname: Nesbitt
authors.forename: M L
authors.initials: ML
authors.lastname: Scott
mesh_terms: D000818:Animals; D000906:Antibodies; D001788:Blood Grouping and Crossmatching / Q000379:methods*; D002460:Cell Line; D006801:Humans; D007144:Immunoglobulin J-Chains / Q000494:pharmacology*; D007075:Immunoglobulin M / Q000276:immunology*; D007626:Kell Blood-Group System / Q000276:immunology*; D011994:Recombinant Proteins
chemical_list: D000906:Antibodies; D007144:Immunoglobulin J-Chains; D007075:Immunoglobulin M; D007626:Kell Blood-Group System; D011994:Recombinant Proteins