Biotechnology Letters

, Volume 33, Issue 4, pp 655–661

Optimization of phage-immobilized ELISA for autoantibody profiling in human sera

  • Mi-Kyung Woo
  • Chang-Kyu Heo
  • Hae-Min Hwang
  • Jeong-Heon Ko
  • Hyang-Sook Yoo
  • Eun-Wie Cho
Original Research Paper

Abstract

Phage libraries displaying cDNA or random peptides have been used for profiling autoantibodies in cancer. The detection of autoantibodies in human sera using phages displaying specific epitopes is usually performed by phage-immobilized ELISAs which can detect specific antibodies without identification of whole antigens. However, these ELISAs can give feeble detection signals that are indistinguishable from background signals which are caused by human sera. To improve the usefulness of phage ELISA for human sera, the conditions for each step in phage ELISA were optimized. The antigenicity of phage antigens was maximal when using coating buffer of neutral pH. By using protein-free blocking buffer and pre-adsorbing human sera with phage host cell ER2738 extracts significantly decreased non-specific signals. Finally, when these conditions were applied to phage ELISA using K10P1, the values of the negative controls were concentrated near cutoff values, which made the assay more reliable. The optimized phage ELISA conditions described here would increase the efficacy of detection specific autoantibodies in human sera.

Keywords

Blocking buffer Coating buffer Human serum Phage ELISA Pre-adsorption 

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Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Mi-Kyung Woo
    • 1
  • Chang-Kyu Heo
    • 1
  • Hae-Min Hwang
    • 1
  • Jeong-Heon Ko
    • 1
  • Hyang-Sook Yoo
    • 1
  • Eun-Wie Cho
    • 1
  1. 1.Daejeon-KRIBB-FHCRC Cooperation Research CenterKorea Research Institute of Bioscience and Biotechnology (KRIBB)DaejeonSouth Korea

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