[Improvement in affinity of phage antibodies against HBsAg by light chain shuffling].
OBJECTIVE
We had cloned an Fd gene of human antibody against HBsAg previously. The objective of this experiment is to improve the affinity phage antibody by chain-shuffling.


METHODS
Human antibody light chain gene repertoire was generated by RT-PCR from human peripheral blood lymphocyte, and then phage antibody sublibrary was constructed by inserting the repertoire into the phagmid which contained the Fd gene. The matched gene of light chain was selected by biopaning.


RESULTS
After three rounds of selection, eight clones with higher absorbance than that of original clone at 490 nm in ELISA were obtained. DNA sequencing showed three of the five VL genes were kappa type, and the other two were lambda type.


CONCLUSIONS
The results indicated that the affinity of chain shuffled phage antibodies(phab) were improved. The specificity of the selected phab was also confirmed.
Mao
C
C
Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing.
Song
H
H
Li
J
J
D000915:Antibody Affinity; D000918:Antibody Specificity; D001435:Bacteriophages / Q000235:genetics*; D001483:Base Sequence; D006510:Hepatitis B Antibodies / Q000235:genetics* / Q000276:immunology; D006514:Hepatitis B Surface Antigens / Q000276:immunology*; D006801:Humans; D007147:Immunoglobulin Light Chains / Q000235:genetics; D008969:Molecular Sequence Data; D020133:Reverse Transcriptase Polymerase Chain Reaction
D006510:Hepatitis B Antibodies; D006514:Hepatitis B Surface Antigens; D007147:Immunoglobulin Light Chains