Identification of two aberrant transcripts derived from a hybridoma with amplification of functional immunoglobulin variable genes.
Murine monoclonal antibodies (mAbs) are widely used but have limitations if administered in humans. The use of chimeric or humanized mAbs can reduce immunogenicity. The first step in producing such mAbs is to clone murine variable genes from a hybridoma, but it is possible to amplify both functional and aberrant variable genes, as they coexist in the hybridoma. During the development of a murine-human chimeric antibody, we have cloned from a hybridoma the functional heavy chain variable region (V(H)) and light chain variable region (V(L)) genes of a mAb that blocks the binding of anthrax lethal factor to protective antigen. In this study, we report the detection of two aberrant transcripts from a hybridoma produced using myeloma cell line OUR-1, the development of a method to distinguish between the functional and abundant aberrant V(L) transcripts, and the origins of these aberrant genes. The aberrant V(L) gene is derived from OUR-1 cells, while the aberrant V(H) gene might derive from antibody repertoires in B cells or from gene rearrangement in the hybridoma cells. The aberrant V(H) and V(L) genes in this study may facilitate discrimination between the functional and aberrant variable genes from hybridoma cells.
Ding
Guipeng
G
Department of Pathology, Nanjing Medical University, Nanjing, China.
Chen
Ximin
X
Zhu
Jin
J
Cao
Brian
B
D000818:Animals; D001483:Base Sequence; D045744:Cell Line, Tumor; D005784:Gene Amplification; D006801:Humans; D006825:Hybridomas; D007135:Immunoglobulin Variable Region / Q000235:genetics* / Q000276:immunology*; D051379:Mice; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D014158:Transcription, Genetic
D007135:Immunoglobulin Variable Region