Generation of a large complex antibody library from multiple donors.
We have generated a large complex library of single chain antibodies based on four individual libraries from each of 50 donors. DNA coding for the heavy and light chain variable domains of the IgM and IgG repertoires was amplified by PCR using two different sets of primers. Each individual library was composed of approximately 1-5x10(7) independent clones giving a final combined library of 4x10(9) members. Screening this library by phage display of single chain antibodies with small haptens, peptides and proteins yielded specific antibodies for each class of antigen.
Little
M
M
German Cancer Research Center, Recombinant Antibody Unit (D0500), Diagnostics and Experimental Therapy Programme, INF 280, 69120, Heidelberg, Germany. m.little@dkfz-heidelberg.de
Welschof
M
M
Braunagel
M
M
Hermes
I
I
Christ
C
C
Keller
A
A
Rohrbach
P
P
Kürschner
T
T
Schmidt
S
S
Kleist
C
C
Terness
P
P
D001782:Blood Donors; D015723:Gene Library; D005803:Genes, Immunoglobulin; D006801:Humans; D007074:Immunoglobulin G / Q000235:genetics; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D007147:Immunoglobulin Light Chains / Q000235:genetics*; D007075:Immunoglobulin M / Q000235:genetics; D007135:Immunoglobulin Variable Region / Q000235:genetics*; D019151:Peptide Library; D016133:Polymerase Chain Reaction
D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007075:Immunoglobulin M; D007135:Immunoglobulin Variable Region; D019151:Peptide Library