[Epitope identification of monoclonal antibody 5C6 against 14-3-3 protein of Schistosoma japonicum].

Abstract

OBJECTIVE:

To identify the epitope of monoclonal antibody (McAb) 5C6 against 14-3-3 protein of Schistosoma japonicum by phage display peptide library.

METHODS:

The phage display 12-peptide library was screened with purified McAb 5C6 against 14-3-3 protein of S. japonicum three rounds of bio-panning "adsorption-elution-amplification" to enrich the specific binding phages. The single phage clones selected randomly were amplified, their genomic DNA were extracted and sequenced. The immune response characterization of phages with the same or high homologous foreign inserted DNA sequence was identified by ELISA and Western blotting for further defining the epitope recognized by McAb 5C6.

RESULTS:

A total of 33 single phage clones were selected and sequenced. Among them, 25 shared the same foreign inserted DNA sequence of 5'-CCACCTAGTAGCAGACCGATTCTCAGTCGAAGGAAA-3', encoding a deduced peptide PPSSRPILSRRK. This peptide was not homologue to Sj14-3-3 protein or any other known native protein in the world. The results of Western blotting showed that this peptide could be recognized by the sera of patients with schistosomiasis, but not by those of healthy persons.

CONCLUSION:

The mimic antigen epitope of McAb 5C6 against 14-3-3 protein of S. japonicum, which is a conformational epitope, has been defined successfully in this study.

• Authors: Chun-Yan Qian, Li-Jun Song, Chuan-Xin Yu, Xu-Ren Yin, Jie Wang, Wei Zhang, Yi Jin, Xue-Dan Ke
• Journal: Zhongguo xue xi chong bing fang zhi za zhi = Chinese journal of schistosomiasis control (Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi) Vol. 24 Issue 1 Pg. 45-9 (Feb 2012) ISSN: 1005-6661 [Print] China
• PMID: 22590863 (Publication Type: English Abstract, Journal Article)

Chemical References

• 14-3-3 Proteins
• Antibodies, Monoclonal
• Epitopes
• Peptide Library

Topics

• 14-3-3 Proteins (genetics, immunology)
• Animals
• Antibodies, Monoclonal (genetics, immunology)
• Antigen-Antibody Reactions
• Base Sequence
• Epitopes (chemistry, genetics, immunology)
• Humans
• Peptide Library
• Schistosoma japonicum (immunology, isolation & purification)