TY - JOUR
T1 - Forced expression of AID facilitates the isolation of class switch variants from hybridoma cells
AU - Iglesias-Ussel, Maria D.
AU - Fan, Manxia
AU - Li, Ziqiang
AU - Martin, Alberto
AU - Scharff, Matthew D.
N1 - Funding Information:
We would like to thank P. Y. Yuan for technical assistance. This work was supported by grants from the National Institutes of Health to M. D. Scharff (CA 72649, CA102705, and AI 43937), who is also supported by the Harry Eagle Chair provided by the National Women's Division of the Albert Einstein College of Medicine. M.D. Iglesias-Ussel was a fellow of the Ministerio de Educación, Cultura y Deporte (Spain) and is currently supported by a Fellowship from the Northeast Biodefense Center (AI57158). Z. L. was supported by a Cancer Research Institute Postdoctoral Fellowship and is currently a Special Fellow of The Leukemia and Lymphoma Society. A. Martin was a special fellow of the Leukemia and Lymphoma Society and currently is a Canada Research Chair in the Department of Immunology at the University of Toronto.
PY - 2006/10/30
Y1 - 2006/10/30
N2 - Monoclonal antibodies are used in the treatment and diagnosis of diseases and to study the protective and adverse functions of antibodies in vitro and in vivo. Since the isotype determines the effector function, half-life in the serum and distribution throughout the body, it would be useful to have a battery of antibodies with the same binding site associated with different isotypes. However, since hybridomas switch isotypes at very low frequencies in tissue culture, it has been difficult and very labor intensive to isolate panels of class switch variants. We show here that stable transfection of activation-induced cytidine deaminase (AID) in hybridomas increased their frequency of switching to a level that greatly facilitated the isolation of subclones expressing monoclonal antibodies of different isotypes. Although forced expression of AID also increased the frequency of somatic hypermutation in the immunoglobulin variable regions that encode the antigen binding site, antigen recognition was retained in the isotype switched antibodies.
AB - Monoclonal antibodies are used in the treatment and diagnosis of diseases and to study the protective and adverse functions of antibodies in vitro and in vivo. Since the isotype determines the effector function, half-life in the serum and distribution throughout the body, it would be useful to have a battery of antibodies with the same binding site associated with different isotypes. However, since hybridomas switch isotypes at very low frequencies in tissue culture, it has been difficult and very labor intensive to isolate panels of class switch variants. We show here that stable transfection of activation-induced cytidine deaminase (AID) in hybridomas increased their frequency of switching to a level that greatly facilitated the isolation of subclones expressing monoclonal antibodies of different isotypes. Although forced expression of AID also increased the frequency of somatic hypermutation in the immunoglobulin variable regions that encode the antigen binding site, antigen recognition was retained in the isotype switched antibodies.
KW - AID
KW - CSR
KW - hybridoma
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U2 - 10.1016/j.jim.2006.08.002
DO - 10.1016/j.jim.2006.08.002
M3 - Article
C2 - 16997317
AN - SCOPUS:33750030491
SN - 0022-1759
VL - 316
SP - 59
EP - 66
JO - Journal of Immunological Methods
JF - Journal of Immunological Methods
IS - 1-2
ER -