title: Rapid specific amplification of rat antibody cDNA from nine hybridomas in the presence of myeloma light chains.
abstract: Most monoclonal antibodies to mouse antigens have been derived from rat spleen-mouse myeloma fusions. Many resultant hybridomas express one of several myeloma kappa chain transcripts, even though the parent myeloma may have been ascribed as not expressing light chain protein. Previous reports have only differentiated against one of these mouse light chains. We have found at least three different myeloma kappa transcripts in the panel of nine hybridomas that were derived from four different myeloma parents. We have designed an amplification strategy that differentiates the rearranged rat kappa chain from all mouse light chains. Moreover, this method is expedient as it requires minimal downstream manipulation.
authors.affiliation: The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia.
authors.forename: Jamie L
authors.initials: JL
authors.lastname: Brady
authors.forename: Alexandra J
authors.initials: AJ
authors.lastname: Corbett
authors.forename: Brent S
authors.initials: BS
authors.lastname: McKenzie
authors.forename: Andrew M
authors.initials: AM
authors.lastname: Lew
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D003001:Cloning, Molecular; D006224:Cricetinae; D017931:DNA Primers; D018076:DNA, Complementary / Q000378:metabolism*; D004274:DNA, Recombinant / Q000378:metabolism*; D006825:Hybridomas / Q000276:immunology; D007147:Immunoglobulin Light Chains / Q000378:metabolism; D007145:Immunoglobulin kappa-Chains / Q000235:genetics* / Q000378:metabolism; D007136:Immunoglobulins / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D009194:Myeloma Proteins / Q000235:genetics* / Q000378:metabolism; D016133:Polymerase Chain Reaction / Q000379:methods*; D051381:Rats; D017386:Sequence Homology, Amino Acid
chemical_list: D017931:DNA Primers; D018076:DNA, Complementary; D004274:DNA, Recombinant; D007147:Immunoglobulin Light Chains; D007145:Immunoglobulin kappa-Chains; D007136:Immunoglobulins; D009194:Myeloma Proteins; C017551:myeloma immunoglobulins