7 research outputs found
Rapid Isolation of Immunoglobulin Variable Genes from Cell Lysates of Rat Hybridomas by Polymerase Chain Reaction
Cloning of cDNAs Encoding the Variable Domains of Antibody BrE-3 and Construction of a Chimeric Antibody
Immunoglobulin V Regions of a Bactericidal Anti-Neisseria meningitidis Outer Membrane Protein Monoclonal Antibody
Polymemse Chain Reaction Using Mixed Primers: Cloning of Human Monoclonal Antibody Variable Region Genes from Single Hybridoma Cells
We describe a general approach to rapidly obtain the DNA sequence encoding the variable region of any immunoglobulin chain using the polymerase chain reaction and a mixture of upstream primers corresponding to the leader sequence, and one downstream primer designed from the conserved nucleotide sequence of the constant region. The approach was applied to five different hybridomas producing human monoclonal antibodies and variable regions for both bold gamma and mu heavy chain and kappa and lambda light chain genes were successfully cloned. cDNA encoding variable regions could be amplified from single hybridoma cells isolated by micromanipulation. This approach will permit analysis of B cell clonal ontogeny, antibody diversity and lymphoma cell progression and heterogeneity. It will also facilitate structural and functional studies of immunoglobulins as well as the rapid construction of chimeric antibodies
The origin of B-chronic lymphocytic leukaemia (B-CLL) and its relationship to hairy cell leukaemia (HCL).
The origin of B-chronic lymphocytic leukaemia and its relationship to hairy cell leukaemia have been investigated by functional and immunophenotipic approache