Schistosoma japonicum: Construction of phage display antibody library and its application in the immunodiagnosis of infection

Dai Xiong Chen, Ai He, Xi Mei Zhan, Mu Hua Yu, Zhi Gang Lei, Jin Xiu Meng, Zhuo Ya Li, Yu Liang, Rui Lin Zhang

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Background. A monoclonal antibody would be an effective tool for the detection of circulating antigens in the serum of patients with schistosomiasis, but the traditional way of producing monoclonal antibodies is not cost-effective. The objective of this study was to find a new method for the large-scale production of monoclonal antibodies against Schistosoma japonicum (Sj). Methods. A phage display antibody library for Sj was constructed. To obtain a single-chain variable fragment antibody (scFv) against Sj, the library was screened with metabolic antigens from adult Sj worms (Sj-MAg) using enzyme-linked immunosorbent assay. The soluble scFvs selected were used to detect Sj antigens in the serum of acute and chronic schistosomiasis patients. Results. Six positive clones with good reactivity to Sj-MAg were obtained from the phage display antibody library of about 1. 07 × 106 individual clones. Only two of these six clones bound specifically to Sj-MAg and were chosen for further analysis. Specific soluble anti-Sj-MAg scFvs were produced by inducing the 2 clones with isopropyl-D-thiogalactopyranoside. The characteristics of the scFvs were then determined. The results of Western blot showed that these scFvs could bind to Sj-MAg specifically and had a molecular weight of about 31 kD. When testing serum from schistosomiasis patients with one of the two specific scFvs, its sensitivity was found to be 60% and 37% in acute and chronic patients, respectively, with a specificity of 90%. When the two specific scFvs were combined, their sensitivity was found to be 75% and 57% in acute and chronic patients, respectively, with a specificity of 85%. Conclusions. The results indicate that the scFvs are potentially useful for the diagnosis of schistosomiasis. The library construction also provides a useful tool for the further screening of other antibodies for both diagnostic and immunotherapeutic applications and for epitope analysis and vaccine design.

Original languageEnglish (US)
Pages (from-to)1697-1703
Number of pages7
JournalChinese Medical Journal
Volume117
Issue number11
StatePublished - Nov 2004
Externally publishedYes

Fingerprint

Schistosoma japonicum
Immunologic Tests
Bacteriophages
Antibodies
Infection
Schistosomiasis
Clone Cells
Monoclonal Antibodies
Antigens
Libraries
Serum
Thiogalactosides
Single-Chain Antibodies
Epitopes
Vaccines
Molecular Weight
Western Blotting
Enzyme-Linked Immunosorbent Assay

Keywords

  • Phage display antibody library
  • Schistosoma japonicum
  • Serodiagnosis
  • Single-chain variable fragment antibody

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Chen, D. X., He, A., Zhan, X. M., Yu, M. H., Lei, Z. G., Meng, J. X., ... Zhang, R. L. (2004). Schistosoma japonicum: Construction of phage display antibody library and its application in the immunodiagnosis of infection. Chinese Medical Journal, 117(11), 1697-1703.

Schistosoma japonicum : Construction of phage display antibody library and its application in the immunodiagnosis of infection. / Chen, Dai Xiong; He, Ai; Zhan, Xi Mei; Yu, Mu Hua; Lei, Zhi Gang; Meng, Jin Xiu; Li, Zhuo Ya; Liang, Yu; Zhang, Rui Lin.

In: Chinese Medical Journal, Vol. 117, No. 11, 11.2004, p. 1697-1703.

Research output: Contribution to journalArticle

Chen, DX, He, A, Zhan, XM, Yu, MH, Lei, ZG, Meng, JX, Li, ZY, Liang, Y & Zhang, RL 2004, 'Schistosoma japonicum: Construction of phage display antibody library and its application in the immunodiagnosis of infection', Chinese Medical Journal, vol. 117, no. 11, pp. 1697-1703.
Chen, Dai Xiong ; He, Ai ; Zhan, Xi Mei ; Yu, Mu Hua ; Lei, Zhi Gang ; Meng, Jin Xiu ; Li, Zhuo Ya ; Liang, Yu ; Zhang, Rui Lin. / Schistosoma japonicum : Construction of phage display antibody library and its application in the immunodiagnosis of infection. In: Chinese Medical Journal. 2004 ; Vol. 117, No. 11. pp. 1697-1703.
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abstract = "Background. A monoclonal antibody would be an effective tool for the detection of circulating antigens in the serum of patients with schistosomiasis, but the traditional way of producing monoclonal antibodies is not cost-effective. The objective of this study was to find a new method for the large-scale production of monoclonal antibodies against Schistosoma japonicum (Sj). Methods. A phage display antibody library for Sj was constructed. To obtain a single-chain variable fragment antibody (scFv) against Sj, the library was screened with metabolic antigens from adult Sj worms (Sj-MAg) using enzyme-linked immunosorbent assay. The soluble scFvs selected were used to detect Sj antigens in the serum of acute and chronic schistosomiasis patients. Results. Six positive clones with good reactivity to Sj-MAg were obtained from the phage display antibody library of about 1. 07 × 106 individual clones. Only two of these six clones bound specifically to Sj-MAg and were chosen for further analysis. Specific soluble anti-Sj-MAg scFvs were produced by inducing the 2 clones with isopropyl-D-thiogalactopyranoside. The characteristics of the scFvs were then determined. The results of Western blot showed that these scFvs could bind to Sj-MAg specifically and had a molecular weight of about 31 kD. When testing serum from schistosomiasis patients with one of the two specific scFvs, its sensitivity was found to be 60{\%} and 37{\%} in acute and chronic patients, respectively, with a specificity of 90{\%}. When the two specific scFvs were combined, their sensitivity was found to be 75{\%} and 57{\%} in acute and chronic patients, respectively, with a specificity of 85{\%}. Conclusions. The results indicate that the scFvs are potentially useful for the diagnosis of schistosomiasis. The library construction also provides a useful tool for the further screening of other antibodies for both diagnostic and immunotherapeutic applications and for epitope analysis and vaccine design.",
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AU - Yu, Mu Hua

AU - Lei, Zhi Gang

AU - Meng, Jin Xiu

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