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The preparation and therapeutic roles of scFv-Fc antibody against Staphylococcus aureus infection to control bovine mastitis

  • Man Wang
  • Tingting Wang
  • Yu Guan
  • Fengqing Wang
  • Jianguo Zhu
Biotechnological products and process engineering

Abstract

Staphylococcus aureus–induced bovine mastitis causes significant losses to the dairy industry and available vaccines do not confer adequate protection. As a more attractive alternative, we propose the use of antibody (Ab) therapy. In our previous study, we constructed a bovine single-chain variable fragment (scFv) Ab phage display and successfully obtained scFvs that bound to S. aureus antigens with high affinity. Here, we describe a novel Ab against S. aureus (scFv-Fc Ab). To construct the scFv-Fc Ab, the scFv Ab was genetically fused to the Fc fragment of a bovine IgG1 Ab. Western blot analysis showed that the bovine scFvs-Fc Abs were successfully expressed with horseradish peroxidase–conjugated goat-anti-bovine IgG (Fc) Ab in Escherichia coli cells. The purified bovine scFvs-Fc Abs had good binding activity to S. aureus and effectively inhibited the bacterial growth in culture medium and bovine scFvs-Fc Abs enhanced phagocytosis of S. aureus by neutrophils isolated from peripheral blood in a dose-dependent manner. In the experiment of bovine scFvs-Fc Abs for the treatment of S. aureus–induced bovine mastitis, the total effective percentage reached 82% (9/11). These novel bovine scFvs-Fc Abs may be useful as therapeutic candidates for the prevention and treatment of S. aureus–induced bovine mastitis.

Keywords

Bovine mastitis Staphylococcus aureus Single-chain variable region fragment (scFv) Fc Phagocytosis 

Notes

Funding

This study was supported by the National Key Research and Development Program of China (Grant Number: 2018YFD0501600).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This study was conducted in accordance with all applicable international, national, and institutional guidelines for the care and use of animals.

Supplementary material

253_2018_9548_MOESM1_ESM.pdf (262 kb)
ESM 1 (PDF 261 kb)

References

  1. Barkema HW, Schukken YH, Zadoks RN (2006) Invited review: the role of cow, pathogen, and treatment regimen in the therapeutic success of bovine Staphylococcus aureus mastitis. J Dairy Sci 89:1877–1895CrossRefGoogle Scholar
  2. Bax HJ, Josephs DH, Pellizzari G, Spicer JF, Montes A, Karagiannis SN (2016) Therapeutic targets and new directions for antibodies developed for ovarian cancer. MAbs 8:1437–1455CrossRefGoogle Scholar
  3. Bird RE, Hardman KD, Jacobson JW, Johnson S, Kaufman BM, Lee SM, Lee T, Pope SH, Riordan GS, Whitlow M (1988) Single-chain antigen-binding proteins. Science 242:423–426CrossRefGoogle Scholar
  4. Bradley AJ (2002) Bovine mastitis: an evolving disease. Vet J 164:116–128CrossRefGoogle Scholar
  5. Brouillette E, Malouin F (2005) The pathogenesis and control of Staphylococcus aureus-induced mastitis: study models in the mouse. Microbes Infect 7:560–568CrossRefGoogle Scholar
  6. Feige MJ, Hendershot LM, Buchner J (2009) How antibodies fold. Trends Biochem Sci 35:189–198CrossRefGoogle Scholar
  7. Furth PA, Shamay A, Wall R, Hennighausen L (1992) Gene transfer into somatic tissues by jet injection. Anal Biochem 205:365–368CrossRefGoogle Scholar
  8. Harris JM, Chess RB (2003) Effect of pegylation on pharmaceuticals. Nat Rev Drug Discov 2:214–221CrossRefGoogle Scholar
  9. Jazayeri JA, Carroll GJ (2008) Fc-based cytokines: prospects for engineering superior therapeutics. Biodrugs 22:11–26CrossRefGoogle Scholar
  10. Kaneko E, Niwa R (2011) Optimizing therapeutic antibody function: progress with Fc domain engineering. BioDrugs 25:1–11CrossRefGoogle Scholar
  11. Kim JH, Sim DW, Park DS, Jung TG, Lee S, Oh T, Ha JR, Seok SH, Seo MD, Kang HC, Kim YP, Won HS (2016) Bacterial production and structure-functional validation of a recombinant antigen-binding fragment (Fab) of an anti-cancer therapeutic antibody targeting epidermal growth factor receptor. Appl Microbiol Biotechnol 100:10521–10529CrossRefGoogle Scholar
  12. Klinman DM, Yamshchikov G, Ishigatsubo Y (1997) Contribution of CpG motifs to the immunogenicity of DNA vaccines. J Immunol 158:3635–3639PubMedGoogle Scholar
  13. Kontermann RE (2009) Strategies to extend plasma half-lives of recombinant antibodies. Biodrugs 23:93–109CrossRefGoogle Scholar
  14. McAuley A, Jacob J, Kolvenbach CG, Westland K, Lee HJ, Brych SR, Rehder D, Kleemann GR, Brems DN, Matsumura M (2008) Contributions of a disulfide bond to the structure, stability, and dimerization of human IgG1 antibody CH3 Domain. Protein Sci 17:95–106CrossRefGoogle Scholar
  15. Middleton JR (2008) Staphylococcus aureus antigens and challenges in vaccine development. Expert Rev Vaccines 7:805–815CrossRefGoogle Scholar
  16. Middleton JR, Hardin D, Steevens B, Randle R, Tyler JW (2004) Use of somatic cell counts and California mastitis test results from individual quarter milk samples to detect subclinical intramammary infection in dairy cattle from a herd with a high bulk tank somatic cell count. J Ame Vet Med Assoc 224:419–423CrossRefGoogle Scholar
  17. Nordhaug ML, Nesse LL, Norcross NL, Gudding R (1994) A field trial with an experimental vaccine against staphylococcus aureus mastitis in callte: 2. Antibody response. J Dairy Sci 77:1276–1284CrossRefGoogle Scholar
  18. Ono K, Kamihira M, Kuga Y, Matsumoto H, Hotta A, Itoh T, Nishijima K, Nakamura N, Matsuda H, Lijima S (2003) Production of anti-prion scFv-Fc fusion proteins by recombinant animal cells. J Biosci Bioeng 95:231–238CrossRefGoogle Scholar
  19. Pennini ME, De Marco A, Pelletier M, Bonnell J, Cvitkovic R, Beltramello M, Cameroni E, Bianchi S, Zatta F, Zhao W, Xiao XD, Camara MM, DiGiandomenico A, Semenova E, Lanzavecchia A, Warrener P, Suzich J, Wang Q, Corti D, Stover CK (2017) Immune stealth-driven O2 serotype prevalence and potential for therapeutic antibodies against multidrug resistant Klebsiella pneumonia. Nat Commun 8:1991CrossRefGoogle Scholar
  20. Rasetti-Escargueil C, Avril A, Chahboun S, Tierney R, Bak N, Miethe S, Mazuet C, Popoff MR, Thulier P, Hust M, Pelat T, Sesardic D (2015) Development of human-like scFv-Fc antibodies neutralizing Botulinum toxin serotype B. MAbs 7:1161–1177CrossRefGoogle Scholar
  21. Roth JA, Kaeberle ML (1981) Evaluation of bovine polymorphonuclear leukocyte function. Vet Immunol Immunopathol 2:157–174CrossRefGoogle Scholar
  22. Shinefield HR, Black S (2005) Prevention of Staphylococcus aureus infections: advances in vaccine development. Expert Rev Vaccines 4:669–676CrossRefGoogle Scholar
  23. Sokolowska-Wedzina A, Chodaczek G, Chudzian J, Borek A, Zakrzewska M, Otlewski J (2017) High-affinity internalizing human scFv-Fc antibody for targeting FGFR1-overexpressing lung cancer. Mol Cancer Res 15:1040–1050CrossRefGoogle Scholar
  24. Stevens MG, Kehrli ME Jr, Canning PC (1991) A colorimetric assay for quantitating bovine neutrophil bactericidal activity. Vet Immunol Immunopathol 28:45–56CrossRefGoogle Scholar
  25. Symons DBA, Clarkson CA, Beale D (1989) Structure of bovine immunlglobulin constant region heavy chain gamma 1 and gamma 2 genes. Mol Immunol 26:841–850CrossRefGoogle Scholar
  26. Tan GY, Bai LQ, Zhong JJ (2013) Exogenous 1,4-butyrolactone stimulates A-factor-like cascade and validamycin biosynthesis in Streptomyces hygroscopicus 5008. Biotechnol Bioeng 110:2984–2993CrossRefGoogle Scholar
  27. Wang M, Zhang Y, Li BQ, Zhu JG (2015) Construction of scFv that bind both fibronectin-binding protein A and clumping factor A of Stapylococcus aureus. Res Vet Sci 100:109–114CrossRefGoogle Scholar
  28. Wang M, Zhang Y, Zhu JG (2016) Anti-Staphylococcus aureus single-chain variable region fragments provide protection against mastitis in mice. Appl Microbiol Biotechnol 100:2153–2162CrossRefGoogle Scholar
  29. Wolff JA, Malone RW, Williams P, Chong W, Acsadi G, Jani A, Felgner PL (1990) Direct gene transfer into mouse muscle in vivo. Science 247:1465–1468CrossRefGoogle Scholar
  30. Yokota T, Milenic DE, Whitlow M, Schlom J (1992) Rapid tumor penetration of a single-chain Fv and comparison with other immunoglobulin forms. Cancer Res 52:3402–3408PubMedGoogle Scholar
  31. Zhen YH, Jin LJ, Guo J, Li XY, Li Z, Fang R, Xu YP (2008) Characterization of specific egg yolk immunoglobulin (IgY) against mastitis-causing staphylococcus aureus. J Appl Microbiol 105:1529–1535CrossRefGoogle Scholar
  32. Zwick MB, Wang M, Poignard P, Stienler G, Katinger H, Burton DR, Parren PW (2001) Neutralization synergy of human immunodeficiency virus type 1 primary isolates by cocktails of broadly neutralizing antibodies. J Virol 75:12198–12208CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Man Wang
    • 1
  • Tingting Wang
    • 1
  • Yu Guan
    • 1
  • Fengqing Wang
    • 1
  • Jianguo Zhu
    • 1
  1. 1.Shanghai Key Laboratory of Veterinary Biotechnology, School of Agriculture and BiologyShanghai JiaoTong UniversityShanghaiChina

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