Preferential generation of monoclonal IgG-producing hybridomas by use of vesicular stomatitis virus-mediated cell fusion.
Vesicular stomatitis virus (VSV)-mediated cell fusion was recently proposed as an alternative fusion technique to generate monoclonal antibody (MAb)-producing hybridomas. In order to further examine this technique, we made direct comparative experiments among VSV, Sendai virus (SV) and polyethylene glycol (PEG)-mediated cell fusion in generating MAb-producing hybridomas. The distribution of immunoglobulin (Ig) isotypes secreted by the hybridomas obtained, as well as hybridoma yield and specific hybridoma yield, was compared. The results show that VSV-fusion yielded almost the same number of specific hybridomas as SV- and PEG-fusion in spite of its lower fusion frequency. In addition, VSV-fusion preferentially gave Ig-producing, especially IgG-producing, hybridomas. SV-fusion yielded both hybridomas and specific hybridomas with similar frequency to PEG-fusion, but IgM-producers predominated. These results demonstrate that fusion method has a considerable influence on the isotypes of obtained antibodies, and also suggest an advantage of VSV-fusion for production of IgG monoclonal antibodies.
Nagata
S
S
Department of Pathology, National Institute of Health, Tokyo, Japan.
Yamamoto
K
K
Ueno
Y
Y
Kurata
T
T
Chiba
J
J
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D002459:Cell Fusion; D005260:Female; D006825:Hybridomas / Q000276:immunology*; D007074:Immunoglobulin G / Q000096:biosynthesis; D007132:Immunoglobulin Isotypes / Q000096:biosynthesis; D007075:Immunoglobulin M / Q000096:biosynthesis; D051379:Mice; D008807:Mice, Inbred BALB C; D010222:Parainfluenza Virus 1, Human / Q000276:immunology; D011092:Polyethylene Glycols; D014721:Vesicular stomatitis Indiana virus / Q000276:immunology*
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D007132:Immunoglobulin Isotypes; D007075:Immunoglobulin M; D011092:Polyethylene Glycols