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Functional expression of anti-hepatitis B virus (HBV) preS2 antigen scFv by cspA promoter system in Escherichia coli and application as a recognition molecule for single-walled carbon nanotube (SWNT) field effect transistor (FET)

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Abstract

The preS2 antigens of hepatitis B virus (HBV), which causes a serious health problem in the world, have been implicated in hepatocyte cell binding and viral penetration. Therefore, the importance of antibody production against preS2 antigen for early diagnosis of HBV has been well established. In this study, the recombinant HBV preS2 single chain variable fragment (scFv) antibody was successfully expressed in E. coli with the novel cold shock vector (pCold) under the cspA promoter, and its expression level was compared with the pET vector under the T7 promoter. Additionally, a host with an oxidizing cytoplasm, E. coli trxB/gor double mutant, was used to improve the soluble expression. The anti-HBV preS2 scFv using pCold vector was successfully expressed in a soluble and functional form in both wild type and double mutant E. coli, while the scFv using the pET vector was expressed in an insoluble form in spite of using a double mutant providing an oxidizing environment. The induction with 0.05 mM IPTG showed a 2-fold higher functional expression compared to induction with 1 mM IPTG, and the functional expression at the induction temperature (15°C), which is optimal temperature for pCold vector, was improved 2-fold and 3- fold at 4 and 25°C, respectively. The efficacy of anti-HBV preS2 scFv for detecting HBV preS2 antigen was tested and verified by using Ni-decorated single-walled carbon nanotube (SWNT) field effect transistors.

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Authors and Affiliations

  1. Department of Chemical Engineering, Kwangwoon University, Seoul, 139-701, Korea

    Dong Hyun Nam & Yong Hwan Kim

  2. Korea Institute of Science and Technology (KIST), Seoul, 136-791, Korea

    Sun-Ae Jun & Byoung-In Sang

  3. Department of Physics, Chonbuk National University, Chonju, 561-756, Korea

    Young-Seob Lo & Ju-Jin Kim

  4. Advanced Materials Div. Korea Research Institute of Chemical Technology, Daejeon, 305-343, Korea

    Hyunju Chang & Jeong-O Lee

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  1. Sun-Ae Jun
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  2. Dong Hyun Nam
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  3. Young-Seob Lo
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  4. Ju-Jin Kim
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  5. Hyunju Chang
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  6. Jeong-O Lee
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  7. Yong Hwan Kim
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  8. Byoung-In Sang
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Correspondence to Yong Hwan Kim or Byoung-In Sang.

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Jun, SA., Nam, D.H., Lo, YS. et al. Functional expression of anti-hepatitis B virus (HBV) preS2 antigen scFv by cspA promoter system in Escherichia coli and application as a recognition molecule for single-walled carbon nanotube (SWNT) field effect transistor (FET). Biotechnol Bioproc E 15, 810–816 (2010). https://doi.org/10.1007/s12257-009-3040-1

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  • DOI: https://doi.org/10.1007/s12257-009-3040-1

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