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Title 

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

Authors 

Mi-Kyung WooChang Gyu HeoHai Min HwangJeong Heon KoHyang Sook YooEun Wie Cho

Publisher 

Springer Verlag (Germany)

Issue Date 

2011

Citation 

Biotechnology Letters, vol. 33, no. 4, pp. 655-661

Keywords 

Blocking bufferCoating bufferHuman serumPhage ELISAPre-adsorption

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.

ISSN 

0141-5492

Link 

http://dx.doi.org/10.1007/s10529-010-0483-6

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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