[Construction of human combinatorial immunoglobulin library containing HBsAb on the surface of phage].
OBJECTIVE
To construct a human recombinant immunoglobulin library containing HBsAb.


METHODS
The mRNA isolated from peripheral blood lymphocytes of two blood donors with HBsAb were reversely transcribed to the first strand cDNA with oligo-(dT) primer. Human immunoglobulin kappa light chains and heavy chains Fd genes were amplified respectively by PCR with designed oligo nucleotide primers and the first strand cDNA templates. The kappa light chain genes were first cloned into PComb3Hss vector to construct a human recombinant light chain library. The heavy chain genes were subsequently inserted into the corresponding sites of kappa-PComb3Hss plasmid to generate a combinatorial kappa-Fd-PComb3Hss plasmid. The plasmid transformed into E.Coli.XLI-Blue and the E.Coli.XLI-Blue infected by help phage.


RESULTS
The constructed light chains library size was 1.5 x 10(5), and the recombinant frequency of kappa chain genes was 57% and the human combinatorial Fab immunoglobulin library was 5 x 10(5) members and the recombinant frequency of Fab genes was 50%. Finally, the titer of phages in the phage antibody library containing HBsAb was 5 x 10(14) CFU/ml.


CONCLUSIONS
Our work in constructing human immunoglobulin combinatorial library containing HBsAb will be beneficial to further study on selecting Fab phage antibodies of HBsAg from the library.
Wang
Z
Z
Institute for Viral Hepatitis, Chongqing University of Medical Sciences.
Liu
Q
Q
Wan
Z
Z
D006510:Hepatitis B Antibodies / Q000235:genetics*; D006514:Hepatitis B Surface Antigens / Q000276:immunology*; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics; D007145:Immunoglobulin kappa-Chains / Q000235:genetics; D019151:Peptide Library
D006510:Hepatitis B Antibodies; D006514:Hepatitis B Surface Antigens; D007143:Immunoglobulin Heavy Chains; D007145:Immunoglobulin kappa-Chains; D019151:Peptide Library