[Screening of human scFv against IL-8 from phage antibody library]

Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2005 Mar;21(2):185-7.
[Article in Chinese]

Abstract

Aim: 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.

Conclusion: 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.

MeSH terms

  • Antibodies / genetics
  • Antibodies / immunology*
  • Antibodies / isolation & purification*
  • Bacteriophages / genetics*
  • Base Sequence
  • Electrophoresis, Polyacrylamide Gel
  • Escherichia coli / genetics
  • Humans
  • Immunoglobulin Heavy Chains / genetics
  • Immunoglobulin Heavy Chains / immunology
  • Immunoglobulin Variable Region / genetics
  • Immunoglobulin Variable Region / immunology*
  • Interleukin-8 / biosynthesis
  • Interleukin-8 / genetics
  • Interleukin-8 / immunology*
  • Molecular Sequence Data
  • Peptide Library*
  • Psoriasis / immunology
  • Psoriasis / therapy
  • Recombinant Fusion Proteins / biosynthesis
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology
  • Recombinant Fusion Proteins / isolation & purification

Substances

  • Antibodies
  • Immunoglobulin Heavy Chains
  • Immunoglobulin Variable Region
  • Interleukin-8
  • Peptide Library
  • Recombinant Fusion Proteins