[Screening of human scFv against IL-8 from phage antibody library].
OBJECTIVE
To screen human scFv against IL-8 from phage antibody library.


METHODS
IL-8-His fusion protein was expressed in E.coli BL21 (DE3) transfected with prokaryotic expression vector pRSET-IL-8 and was purified by affinity chromatography. Specific antibody was screened by 3 rounds of panning of phage antibody library with the fusion protein. The antigen binding activity and DNA sequences of positive clones were determined and analyzed.


RESULTS
After 3 rounds of panning, 2 positive clones were obtained which could bind IL-8 specifically. DNA sequence analysis showed both of the 2 VH genes belonged to human IgG VH3 subgroup, and the Vlambda gene belonged to human VlambdaDPL5 and VlambdaDPL2 subgroups,respectively.


CONCLUSIONS
It is feasible to obtain human anti-IL-8 antibody by phage display technique, which provides the basis for further research on psoriasis and other related diseases.
Wang
Lei
L
Department of Dermatology, Xijing Hospital, Fourth Military Medical University, Xi'an 710032, China.
Qiao
Yuan-yuan
YY
Wang
Gang
G
Chen
Yu-ping
YP
Yang
Jun
J
Wang
Yan
Y
Liu
Yu-feng
YF
D000906:Antibodies / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification*; D001435:Bacteriophages / Q000235:genetics*; D001483:Base Sequence; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000235:genetics / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000235:genetics / Q000276:immunology*; D016209:Interleukin-8 / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D019151:Peptide Library; D011565:Psoriasis / Q000276:immunology / Q000628:therapy; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification
D000906:Antibodies; D007143:Immunoglobulin Heavy Chains; D007135:Immunoglobulin Variable Region; D016209:Interleukin-8; D019151:Peptide Library; D011993:Recombinant Fusion Proteins