지원사업
학술연구/단체지원/교육 등 연구자 활동을 지속하도록 DBpia가 지원하고 있어요.
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연구자들이 자신의 연구와 전문성을 널리 알리고, 새로운 협력의 기회를 만들 수 있는 네트워킹 공간이에요.
논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 발행연도
- 2014.1
- 수록면
- 124 - 132 (9page)
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Background: Without appropriate culture systems for hepatitis E virus (HEV), sufficient natural viral proteins are difficult to generate for use in serological tests. Therefore, it is im- portant to produce large amounts of HEV recombinant proteins in an economical way. The present study developed ELISAs using 2 truncated forms of the HEV open reading frame (ORF) 2 protein in order to detect anti-HEV IgG in serum samples.
Methods: Two truncated forms of the ORF2 protein were expressed in Escherichia coli and were purified by Ni 2+ -chelate-affinity chromatography (Qiagen, Germany). Two ELISAs were developed using these proteins and were compared with DIA.PRO HEV IgG ELISA kit (DIA.PRO. Italy) in 220 serum samples.
Results: High yields of the target proteins were obtained through codon optimization. The concentration and purity of the proteins were improved with Amicon filters (EMD Millipore, USA). Sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) and West- ern blotting analysis of the resultant proteins showed a protein band of approximately 60 kDa corresponding to ORF2.1 (amino acids 112-660) and a protein band of approxi- mately 55 kDa corresponding to ORF2.2 (amino acids 112-607). Positive agreement, negative agreement, and concordance of the 2 in-house ELISAs compared with DIA.PRO HEV IgG ELISA kit were 87%, 99.5%, and 98.1%, respectively (kappa=0.899, P =0.625).
Conclusions: The newly developed ELISAs are useful for detecting anti-HEV IgG in serum samples and are highly concordant with DIA.PRO HEV IgG ELISA kit.
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참고문헌 (28)
참고문헌 신청
Jacobs D / 1999 / Evaluation of recombinant dense granule antigen 7 (GRA7) of Toxoplasma gondii for detection of immunoglobulin G antibodies and analysis of a major antigenic domain / Clin Diagn Lab Immunol 6:24~29
Liu XS / 2011 / Cloning, codon-optimized expression and homology modeling of structural protein VP1 from foot and mouth disease virus / Afr J Microbiol Res 5:486~495
Wang X / 2010 / Codon optimization enhances secretory expression of Pseudomonas aeruginosa exotoxin A in E. coli / Protein Expr Purif 72:101~106
Garmory HS / 2003 / DNA vaccines: improving expression of antigens / Genet Vaccines Ther 1:2
Ramakrishna L / 2004 / Codon optimization of the tat antigen of human immunodeficiency virus type 1 generates strong immune responses in mice following genetic immunization / J Virol 78:9174~9189
Liu XS / 2011 / Cloning, codon-optimized expression and homology modeling of structural protein VP1 from foot and mouth disease virus / Afr J Microbiol Res 5:486~495
Wang X / 2010 / Codon optimization enhances secretory expression of Pseudomonas aeruginosa exotoxin A in E. coli / Protein Expr Purif 72:101~106
Garmory HS / 2003 / DNA vaccines: improving expression of antigens / Genet Vaccines Ther 1:2
Ramakrishna L / 2004 / Codon optimization of the tat antigen of human immunodeficiency virus type 1 generates strong immune responses in mice following genetic immunization / J Virol 78:9174~9189
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