Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지
DOI QR코드

DOI QR Code

Fusion Peptide Improves Stability and Bioactivity of Single Chain Antibody against Rabies Virus

  • Xi, Hualong (National Engineering Laboratory for AIDS Vaccine, Jilin University) ;
  • Zhang, Kaixin (National Engineering Laboratory for AIDS Vaccine, Jilin University) ;
  • Yin, Yanchun (National Engineering Laboratory for AIDS Vaccine, Jilin University) ;
  • Gu, Tiejun (National Engineering Laboratory for AIDS Vaccine, Jilin University) ;
  • Sun, Qing (National Engineering Laboratory for AIDS Vaccine, Jilin University) ;
  • Shi, Linqing (National Engineering Laboratory for AIDS Vaccine, Jilin University) ;
  • Zhang, Renxia (National Engineering Laboratory for AIDS Vaccine, Jilin University) ;
  • Jiang, Chunlai (National Engineering Laboratory for AIDS Vaccine, Jilin University) ;
  • Kong, Wei (National Engineering Laboratory for AIDS Vaccine, Jilin University) ;
  • Wu, Yongge (National Engineering Laboratory for AIDS Vaccine, Jilin University)
  • Received : 2016.11.23
  • Accepted : 2017.01.05
  • Published : 2017.04.28
Download PDF
⟨ Previous Next ⟩

Abstract

The combination of rabies immunoglobulin (RIG) with a vaccine is currently effective against rabies infections, but improvements are needed. Genetic engineering antibody technology is an attractive approach for developing novel antibodies to replace RIG. In our previous study, a single-chain variable fragment, scFv57R, against rabies virus glycoprotein was constructed. However, its inherent weak stability and short half-life compared with the parent RIG may limit its diagnostic and therapeutic application. Therefore, an acidic tail of synuclein (ATS) derived from the C-terminal acidic tail of human alpha-synuclein protein was fused to the C-terminus of scFv57R in order to help it resist adverse stress and improve the stability and half-life. The tail showed no apparent effect on the preparation procedure and affinity of the protein, nor did it change the neutralizing potency in vitro. In the ELISA test of molecular stability, the ATS fusion form of the protein, scFv57R-ATS, showed an increase in thermal stability and longer half-life in serum than scFv57R. The protection against fatal rabies virus challenge improved after fusing the tail to the scFv, which may be attributed to the improved stability. Thus, the ATS fusion approach presented here is easily implemented and can be used as a new strategy to improve the stability and half-life of engineered antibody proteins for practical applications.

Keywords

References

  1. Nagarajan T, Marissen W, Rupprecht C. 2014. Monoclonal antibodies for the prevention of rabies: theory and clinical practice. Antibody Technol. J. 4: 1-12.
  2. Bourhy H, Dautry-Varsat A, Hotez PJ, Salomon J. 2010. Rabies, still neglected after 125 years of vaccination. PLoS Negl. Trop. Dis. 4: e839-e839. https://doi.org/10.1371/journal.pntd.0000839
  3. Knobel DL, Sarah C, Coleman PG, Fèvre EM, Meltzer MI, Miranda MEG, et al. 2005. Re-evaluating the burden of rabies in Africa and Asia. Bull. World Health Organ. 83: 360-368.
  4. Jaap G, Marissen WE, Weldon WC, Michael N, Hanlon CA, Rice AB, et al. 2006. Comparison of an anti-rabies human monoclonal antibody combination with human polyclonal anti-rabies immune globulin. J. Infect. Dis. 193: 796-801. https://doi.org/10.1086/500470
  5. Richard F, Smith TG, Dyer JL, Xianfu W, Michael N, Rupprecht CE. 2013. Current and future tools for global canine rabies elimination. Antiviral Res. 100: 220-225. https://doi.org/10.1016/j.antiviral.2013.07.004
  6. Nelson AL. 2010. Antibody fragments: hope and hype. MAbs 2: 77-83. https://doi.org/10.4161/mabs.2.1.10786
  7. Bird RE, Hardman KD, Jacobson JW, Johnson S, Kaufman BM, Lee SM, et al. 1988. Single-chain antigen-binding proteins. Science 242: 423-426. https://doi.org/10.1126/science.3140379
  8. Weisser NE, Hall JC. 2009. Applications of single-chain variable fragment antibodies in therapeutics and diagnostics. Biotechnol. Adv. 27: 502-520. https://doi.org/10.1016/j.biotechadv.2009.04.004
  9. Cheng Y, Li Z, Xi H, Gu T, Yuan R, Chen X, et al. 2015. A VL-linker-VH orientation dependent single chain variable antibody fragment against rabies virus G protein with enhanced neutralizing potency in vivo. Protein Pept. Lett. 23: 24-32. https://doi.org/10.2174/0929866522666151026122752
  10. Bakker A, Python C, Kissling C, Pandya P, Marissen W, Brink M, et al. 2008. First administration to humans of a monoclonal antibody cocktail against rabies virus: safety, tolerability, and neutralizing activity. Vaccine 26: 5922-5927. https://doi.org/10.1016/j.vaccine.2008.08.050
  11. Myun PS, Keun Jae A, Young JH, Jeon Han P, Jongsun K. 2004. Effects of novel peptides derived from the acidic tail of synuclein (ATS) on the aggregation and stability of fusion proteins. Protein Eng. Des. Sel. 17: 251-260. https://doi.org/10.1093/protein/gzh029
  12. Lee EN, Kim YM, Lee HJ, Sang WP, Han YJ, Lee JM, et al. 2005. Stabilizing peptide fusion for solving the stability and solubility problems of therapeutic proteins. Pharm. Res. 22: 1735-1746. https://doi.org/10.1007/s11095-005-6489-4
  13. Goldman ER, Brozozog-Lee PA, Dan Z, Turner KB, Walper SA, Liu JL, Anderson GP. 2014. Negative tail fusions can improve ruggedness of single domain antibodies. Protein Expr. Purif. 95: 226-232. https://doi.org/10.1016/j.pep.2014.01.003
  14. Duan Y, Gu TJ, Jiang CL, Yuan RS, Zhang HF, Hou HJ, et al. 2012. A novel disulfide-stabilized single-chain variable antibody fragment against rabies virus G protein with enhanced in vivo neutralizing potency. Mol. Immunol. 51: 188-196. https://doi.org/10.1016/j.molimm.2012.03.015
  15. Yuan R, Chen X, Chen Y, Gu T, Xi H, Duan Y, et al. 2014. Preparation and diagnostic use of a novel recombinant single-chain antibody against rabies virus glycoprotein. Appl. Microbiol. Biotechnol. 98: 1547-1555. https://doi.org/10.1007/s00253-013-5351-6
  16. Wu X, Yan X, Yang D. 2001. Affinity maturation of a human single-chain Fv by phage display. Chinese J. Microbiol. Immunol. 21: 156-159.
  17. Reed LJ, Muench H. 1938. A simple method of estimating fifty per cent endpoints. Am. J. Hyg. 27: 493-497.
  18. Duan Y, Gu T, Zhang X, Jiang C, Yuan R, Li Z, et al. 2014. Negative effects of a disulfide bond mismatch in anti-rabies G protein single-chain antibody variable fragment FV57. Mol. Immunol. 59: 136-141. https://doi.org/10.1016/j.molimm.2014.01.006
  19. Safar F, Vahideh A, Asghar T, Kamal V, Shiva Ahdi K, Leila R. 2014. Development trends for generation of single-chain antibody fragments. Immunopharmacol. Immunotoxicol. 36: 1-12. https://doi.org/10.3109/08923973.2013.850506
  20. Cai K, Wang H, Bao S, Shi J, Hou X, Gao X, et al. 2008. Novel human 3-domain disulfide-stabilized antibody fragment against glycoprotein of rabies virus. Microb. Infect. 10: 548-555. https://doi.org/10.1016/j.micinf.2008.02.008
  21. Wang DD, Su MM, Sun Y, Huang SL, Wang J, Yan WQ. 2012. Expression, purification and characterization of a human single-chain Fv antibody fragment fused with the Fc of an IgG1 targeting a rabies antigen in Pichia pastoris. Protein Expr. Purif. 86: 75-81. https://doi.org/10.1016/j.pep.2012.08.015
  22. Liu X, Lin H, Tang Q, Li C, Yang S, Wang Z, et al. 2011. Characterization of a human antibody fragment Fab and its calcium phosphate nanoparticles that inhibit rabies virus infection with vaccine. PLoS One 6: e19848. https://doi.org/10.1371/journal.pone.0019848
  23. Jiang H, Xie Y, Burnette A, Roach J, Giardina SL, Hecht TT, et al. 2013. Purification of clinical-grade disulfide stabilized antibody fragment variable-Pseudomonas exotoxin conjugate (dsFv-PE38) expressed in Escherichia coli. Appl. Microbiol. Biotechnol. 97: 621-632. https://doi.org/10.1007/s00253-012-4319-2
  24. Lee JH, Park DY, Lee KJ, Kim YK, So YK, Ryu JS, et al. 2013. Intracellular reprogramming of expression, glycosylation, and function of a plant-derived antiviral therapeutic monoclonal antibody. PLoS One 8: e68772. https://doi.org/10.1371/journal.pone.0068772
  25. Kim Y, Lee H, Lee J, Kim H, Kim J. 2009. Expression of human interferon alpha-1 with enhanced stability via the tagging system of a stabilizing peptide. Protein Expr. Purif. 63: 140-146. https://doi.org/10.1016/j.pep.2008.09.016
  26. Both L, B anyard A C, V an D C, H orton D L, M a JK, Fooks AR. 2012. Passive immunity in the prevention of rabies. Lancet Infect. Dis. 12: 397-407. https://doi.org/10.1016/S1473-3099(11)70340-1
  27. Koprowski H, van der Scheer J, Black J. 1950. Use of hyperimmune antirabies serum concentrates in experimental rabies. Am. J. Med. 8: 412-420. https://doi.org/10.1016/0002-9343(50)90224-5
  28. Sikes RK, Cleary WF, Koprowski H, Wiktor TJ, Kaplan MM. 1971. Effective protection of monkeys against death from street virus by post-exposure administration of tissue-culture rabies vaccine. Bull. World Health Organ. 45: 1-11.
  29. Turki I, Hammami A, Kharmachi H, Mousli M. 2014. Engineering of a recombinant trivalent single-chain variable fragment antibody directed against rabies virus glycoprotein G with improved neutralizing potency. Mol. Immunol. 57: 66-73. https://doi.org/10.1016/j.molimm.2013.08.009
  30. Todorovska A, Roovers RC, Dolezal O, Kortt AA, Hoogenboom HR, Hudson PJ. 2001. Design and application of diabodies, triabodies and tetrabodies for cancer targeting. J. Immunol. Methods 248: 47-66. https://doi.org/10.1016/S0022-1759(00)00342-2

Cited by

  1. Zika Virus Proteins NS2A and NS4A Are Major Antagonists that Reduce IFN-β Promoter Activity Induced by the MDA5/RIG-I Signaling Pathway vol.29, pp.10, 2019, https://doi.org/10.4014/jmb.1909.09017
  2. Fibrillar form of α-synuclein-specific scFv antibody inhibits α-synuclein seeds induced aggregation and toxicity vol.10, pp.None, 2020, https://doi.org/10.1038/s41598-020-65035-8