Rapid specific amplification of rat antibody cDNA from nine hybridomas in the presence of myeloma light chains.
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.
Brady
Jamie L
JL
The Walter and Eliza Hall Institute of Medical Research, Parkville, Australia.
Corbett
Alexandra J
AJ
McKenzie
Brent S
BS
Lew
Andrew M
AM
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
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