Construction of multiform scFv antibodies using linker peptide

Shihua Wang; Cengjie Zheng; Ying Liu; Huirong Zheng; Zonghua Wang

doi:10.1016/S1673-8527(08)60045-4

Volume 35 Issue 5

May 2008

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Article Navigation > Journal of Genetics and Genomics > 2008 > 35(5): 313-316

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Construction of multiform scFv antibodies using linker peptide

doi: 10.1016/S1673-8527(08)60045-4

a
Key Laboratory of Biopesticide and Chemical Biology, Ministry of Education, Fujian Agriculture and Forestry University, Fuzhou 350002, China
b
College of Life Sciences, Fujian Agriculture and Forestry University, Fuzhou 350002, China

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Corresponding author: E-mail address: wangzh@fjau.edu.cn (Zonghua Wang)
Received Date: 2007-09-25
Accepted Date: 2007-12-21
Rev Recd Date: 2007-12-20

Available Online: 2008-05-20

Publish Date: 2008-05-20

Abstract

Abstract

Multiform single chain variable fragments (scFvs) including different length linker scFvs and bispecific scFv were constructed. The linker lengths of 0, 3, 5, 8, 12, and 15 amino acids between V_H and V_L of antideoxynivalenol (anti-DON) scFv were used to analyze the affinities of scFvs. The affinity constants of these scFvs increased when the linker was lower than 12 amino acids. The affinity constant would not change when the linker was longer than 12 amino acids. Fusion gene of anti-DON scFv and antizearalenone (anti-ZEN) scFv was also constructed through connection by a short peptide linker DNA to express a bispecific scFv. The affinity constants assay showed that the two scFvs of fusion bispecific scFv remained their own affinity compared to their parental scFvs. Competitive direct enzyme linked immunosorbent assay was used to detect DON and ZEN in contaminated wheat (Triticum aestivum L.) samples, and the results indicated that this bispecific scFv was applicable in DON and ZEN detection. This work confirmed that bispecific scFv could be successfully obtained, and might also have an application in diagnosing fungal infection, and breeding transgenic plants.
- multiform scFvs,
- bispecific scFv,
- affinity,
- detection

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References(19)

References

[1]	Choi, G.H., Lee, D.H., Min, W.K. et al. Protein Expr. Purif., 35 (2004),pp. 84-92
[2]	Cochlovius, B., Kipriyanov, S.M., Stassar, M.J. et al. Cure of Burkitt's lymphoma in severe combined immunodeficiency mice by T cells, tetravalent CD3 × CD19 tandem diabody, and CD28 costimulation Cancer Res., 60 (2000),pp. 4336-4341
[3]	Dai, H.P., Cao, H., Zhao, X.Y. et al. Construction and characterization of a novel recombinant single-chain variable fragment antibody against White Spot Syndrome Vrius from shrimp J. Immun. Methods, 279 (2003),pp. 267-275
[4]	Danicke, S., Ueberschar, K.H., Valenta, H. et al. Br. Poult. Sci., 45 (2004),pp. 264-272
[5]	diMenna, M.E., Lauren, D.R., Hardacre, A. Mycopathologia, 139 (1997),pp. 165-173
[6]	Eriksen, G.S., Pettersson, H., Lindberg, J.E. Absorption, metabolism and excretion of 3-acetyl DON in pigs Arch. Tierernahr., 57 (2003),pp. 335-345
[7]	Fox, M., Gray, G., Kavanagh, K. et al. J. Microbiol. Methods, 56 (2004),pp. 221-230
[8]	Goyarts, T., Danicke, S. Toxicol. Lett., 163 (2006),pp. 171-182
[9]	Hart, L.P., Braselton, W.E., Stebbins, T.C. Production of zearalenone and deoxynivalenol in commercial sweet corn Plant Dis., 66 (1982),pp. 1133-1135
[10]	Holliger, P., Wing, M., Pound, J.D. et al. Retargeting serum immunoglobulin with bispecific diabodies Nat. Biotechnol., 15 (1997),pp. 632-636
[11]	Liu, M.T., Ram, B.P., Hart, L.P. et al. Indirect enzyme-linked immunosorbent assay for the mycotoxin zearalenonet Appl. Environ. Microbiol., 50 (1985),pp. 332-336
[12]	Lutzf, M.P., Feichtinger, G., Defago, G. et al. Appl. Environ. Microbiol., 69 (2003),pp. 3077-3084
[13]	Naef, A., Senatore, M., Defago, G. FEMS Microbiol. Ecol., 55 (2006),pp. 211-220
[14]	Wang, S.H., Du, X.Y., Huang, Y.M. et al. Detection of deoxynivalenol (DON) based on a single chain fragment variable (scFv) of the anti-DON antibody FEMS Microbiol. Lett., 272 (2007),pp. 214-219
[15]	Wang, S.H., Zhang, J.B., Zhang, Z.P. et al. Anal. Chem., 78 (2006),pp. 997-1004
[16]	World Health Organization
[17]	Stob, M., Baldwin, R.S., Tuite, J. et al. Isolation of an anabolic, uterotrophic compound from corn infested with Gibberella zeae Nature, 196 (1962),pp. 1318-1319
[18]	Yoshizawa, T., Kohno, H., Ikeda, K. et al. A practical method for measuring deoxynivalenol, nivalenol, and T-2 + HT-2 toxin in foods by an enzyme-linked immunosorbent assay using monoclonal antibodies Biosci. Biotechnol. Biochem., 68 (2004),pp. 2076-2085
[19]	Yuan, Q., Clarke, J.R., Zhou, H.R. et al. Molecular cloning, expression, and characterization of a functional single-chain Fv antibody to the mycotoxin zearalenone Appl. Environ. Microbiol., 63 (1997),pp. 263-269