Forced expression of AID facilitates the isolation of class switch variants from hybridoma cells.
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.
Iglesias-Ussel
Maria D
MD
Department of Cell Biology, Albert Einstein College of Medicine, Bronx, NY 10461, USA.
Fan
Manxia
M
Li
Ziqiang
Z
Martin
Alberto
A
Scharff
Matthew D
MD
D000818:Animals; D015153:Blotting, Western; D003564:Cytidine Deaminase / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D006825:Hybridomas / Q000201:enzymology / Q000276:immunology*; D017578:Immunoglobulin Class Switching / Q000235:genetics / Q000276:immunology*; D007132:Immunoglobulin Isotypes / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008810:Mice, Inbred C57BL; D012313:RNA / Q000737:chemistry / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D027041:Somatic Hypermutation, Immunoglobulin / Q000276:immunology; D014162:Transfection; D000098830:AICDA (Activation-Induced Cytidine Deaminase)
D007132:Immunoglobulin Isotypes; D012313:RNA; D000098830:AICDA (Activation-Induced Cytidine Deaminase); D003564:Cytidine Deaminase