A rapid and quantitative method for the evaluation of V gene usage, specificities and the clonal size of B cell repertoires.
The quantitative simultaneous description of both variable region gene usage and antigen specificity of immunoglobulin repertoires is a major goal in immunology. Current quantitative assays are labor intensive and depend on extensive gene expression cloning prior to screening for antigen specificity. Here we describe an alternative method based on high efficiency single B cell cultures coupled with RT-PCR that can be used for rapid characterization of immunoglobulin gene segment usage, clonal size and antigen specificity. This simplified approach should facilitate the study of antibody repertoires expressed by defined B cell subpopulations, the analysis of immune responses to self and nonself-antigens, the development and screening of synthetic antibodies and the accelerated study and screening of neutralizing antibodies to pathogenic threats.
Department of Medicine, University of Alabama at Birmingham, Birmingham, AL 35294, United States.
A M
AM
Vale
J B
JB
Foote
A
A
Granato
Y
Y
Zhuang
R M S
RM
Pereira
U G
UG
Lopes
M
M
Bellio
P D
PD
Burrows
H W
HW
Schroeder
A
A
Nobrega
D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology / Q000276:immunology*; D002999:Clone Cells / Q000276:immunology*; D003001:Cloning, Molecular / Q000379:methods*; D005260:Female; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D012313:RNA / Q000737:chemistry / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA
D007135:Immunoglobulin Variable Region; D012313:RNA