Multiplex screening for functionally rearranged immunoglobulin variable regions reveals expression of hybridoma-specific aberrant V-genes.
Modification of antibody effector functions is commonly performed by chimerization or humanization. Cloning of antibody variable regions from hybridomas represents a first step that is frequently hampered by the expression of non-functionally rearranged variable regions in hybridoma cells that originate from MOPC21-derived fusion partners. We now present a simple method to clone functionally rearranged V-genes, based on V-gene-specific multiplex PCR screening. Using this method we document the expression of aberrant V-genes that originate from the original B-cell used for the hybridoma generation, not from the fusion partner, and are - thus - hybridoma specific.
Immunotherapy Laboratory, Department of Immunology, Rm KC.02-085.2, University Medical Center Utrecht, Lundlaan 6, 3584 EA, Utrecht, The Netherlands.
G
G
Vidarsson
J G
JG
van de Winkel
M A
MA
van Dijk
D000818:Animals; D001483:Base Sequence; D015322:Gene Rearrangement, B-Lymphocyte; D005803:Genes, Immunoglobulin; D006825:Hybridomas; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D007158:Immunologic Techniques; D051379:Mice; D008969:Molecular Sequence Data
D007135:Immunoglobulin Variable Region