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Synthesis, Characterization and Lipophilicity Study of Brucella abortus’ Immunogenic Peptide Sequence That Can Be Used in the Future Vaccination Studies

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Brucellosis is primarily a zoonotic disease caused by members of the Brucella genus which consists of 11 recognized species based on pathogenicity and host preferences. Peptide vaccines are an alternative strategy to conventional vaccines based on the use of short peptide sequences and engineered to induce highly targeted immune responses, avoiding allergenic and/or reactogenic sequences. In this study, antigenic peptide sequence of Brucella abortus, WLAEIKQRSLMVHG, was chemically synthesized by adding tryptophan to the N-terminus of sequence, purified and characterized for the first time in the literature. Molecular weight of the peptide was determined by using LC–MS. A linear response (R2 > 0.998) is observed for peptide in the range of 2–250 µg/mL. The LOD and LOQ values are 0.08 and 0.27 µg/kg respectively. Precision and accuracy ranges were found to be %RSD < 0.2 and 57.3–103.2%, respectively. Fluorescence property of the peptide was shown via fluorescence spectroscopy measurement. The 3D de novo structure of the peptide was predicted with PEP-FOLD server. According to the data obtained from the lipophilicity study (LogD7.4 = − 3.093 ± 0.195), the peptide can not cross the blood–brain barrier if applied via intravenously. This has shown us that peptide can be used as a peptide candidate at the vaccine studies. The production of antigenic peptides as in this study is the main component of the preparation of synthetic vaccine systems.

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  1. Abubakar M, Mansoor M, Arshed MJ (2012) Bovine brucellosis: old and new concepts with Pakistan perspective. Pak Vet J 32:147–155

  2. Alston RW, Lasagna M, Grimsley GR, Scholtz JM, Reinhart GD, Pace CN (2008) Peptide sequence and conformation strongly influence tryptophan fluorescence. Biophys J 94:2280–2287

  3. Bacsa B, Desai B, Dibo G, Kappe CO (2006) Rapid solid-phase peptide synthesis using thermal and controlled microwave irradiation. J Pept Sci 12:633–638. https://doi.org/10.1002/psc.771

  4. Bhal SK (2007) Lipophilicity descriptors: understanding when to use LogP & LogD ACD/Labs PhysChem Software Application Notes

  5. De BK et al (2008) Novel Brucella strain (BO1) associated with a prosthetic breast implant infection. J Clin Microbiol 46:43–49. https://doi.org/10.1128/JCM.01494-07

  6. Demarque DP, Crotti AE, Vessecchi R, Lopes JL, Lopes NP (2016) Fragmentation reactions using electrospray ionization mass spectrometry: an important tool for the structural elucidation and characterization of synthetic and natural products. Nat Prod Rep 33:432–455

  7. Derman S, Kizilbey K, Mansuroglu B, Mustafaeva Z (2014) Synthesis and characterization of canine parvovirus (CPV) VP2 W-7L-20 synthetic peptide for synthetic vaccine. Fresen Environ Bull 23:558–566

  8. Dorneles EM et al (2015) Immune response of calves vaccinated with Brucella abortus S19 or RB51 and revaccinated with RB. PLoS ONE 10:e0136696

  9. Erdelyi M, Gogoll A (2002) Rapid microwave-assisted solid phase peptide synthesis. Synthesis 11:1592–1596

  10. Filenko A, Demchenko M, Mustafaeva Z, Osada Y, Mustafaev M (2001) Fluorescence study of Cu2+-induced interaction between albumin and anionic polyelectrolytes. Biomacromolecules 2:270–277

  11. Garcia M (2005) The effect of the mobile phase additives on sensitivity in the analysis of peptides and proteins by high-performance liquid chromatography–electrospray mass spectrometry. J Chromatogr B 825:111–123

  12. Hein RJ (2008) LC-MS analysis of related peptides and anions in the positive mode. Iowa State University, Ames

  13. Leo A, Hansch C, Elkins D (1971) Partition coefficients and their uses. Chem Rev 71:525–616. https://doi.org/10.1021/cr60274a001

  14. Li Z et al (2018) Brucella abortus phosphoglyceromutase and dihydrodipicolinate reductase induce Th1 and Th2-related immune responses World. J Microbiol Biotechnol 34:22

  15. Liu H, Zhang J, Sun H, Xu C, Zhu Y, Xie H (2011) The prediction of peptide charge states for electrospray ionization in mass spectrometry. Procedia Environ Sci 8:483–491

  16. Mannhold R, Kubinyi H, Timmerman H (2008) Lipophilicity in drug action and toxicology. vol 4. Wiley, Hoboken

  17. Mansuroglu B, Mustafaeva Z (2012) Characterization of water-soluble conjugates of polyacrylic acid and antigenic peptide of FMDV by size exclusion chromatography with quadruple detection. Mater Sci Eng C 32:112–118. https://doi.org/10.1016/j.msec.2011.10.004

  18. Matsushita T, Hinou H, Kurogochi M, Shimizu H, Nishimura S (2005) Rapid microwave-assisted solid-phase glycopeptide synthesis. Org Lett 7:877–880. https://doi.org/10.1021/ol0474352

  19. Merrifield RB (1963) Solid phase peptide synthesis. I. The synthesis of a tetrapeptide. J Am Chem Soc 85:2149–2154

  20. Moisa AA, Kolesanova EF (2010) Synthetic peptide vaccines. Biochem Suppl Ser B 4:321–332

  21. Mustafaev M, Mustafaeva Z, Deliloglu-Gurhan SI, Aynagoz G, Unver G, Unal N (2004) Bioconjugates of synthetic peptide epitops of foot-and-mouth disease virus with polyelectrolytes and their immunological activity. J Pept Sci 10:291–291

  22. Mustafaev M, Mustafaeva Z, Mansour B (2006) Covalent conjugation of peptide epitops of VP1 protein of foot-and-mouth disease virus (FMDV) with membrane active anionic polyelectrolytes. J Pept Sci 12:239–239

  23. Mustafaeva Z (2017) Synthesis and characterization of antigenic peptide of sheep pox disease. Kimya Problemlеri 3:255–263

  24. Nava-Parada P, Forni G, Knutson KL, Pease LR, Celis E (2007) Peptide vaccine given with a Toll-like receptor agonist is effective for the treatment and prevention of spontaneous breast tumors. Cancer Res 67:1326–1334. https://doi.org/10.1158/0008-5472.CAN-06-3290

  25. Niessen WM, Manini P, Andreoli R (2006) Matrix effects in quantitative pesticide analysis using liquid chromatography-mass spectrometry. Mass Spectrom Rev 25:881–899. https://doi.org/10.1002/mas.20097

  26. Oka Y, Tsuboi A, Oji Y, Kawase I, Sugiyama H (2008) WT1 peptide vaccine for the treatment of cancer. Curr Opin Immunol 20:211–220. https://doi.org/10.1016/j.coi.2008.04.009

  27. Ozdemir ZO, Topuzogulları M, Karabulut E, Akdeste ZM (2009) Characterization and purification of viral peptides synthesized with microwave assisted solid phase method. Int J Nat Eng Sci 3:45–48

  28. Peptide Property Calculator (2017) http://pepcalc.com/ppc.php. Accessed 28 Dec 2017

  29. Roop RM II, Caswell CC (2017) Metals and the biology and virulence of brucella. Springer, New York

  30. Sarikaya M, Enginar H (2012) Radioabeling of L-tyrosine with 131I and Investigation of radiopharmaceutical potantial Afyon Kocatepe University. J Sci 12:1–9

  31. Schelling E, Diguimbaye C, Daoud S, Nicolet J, Boerlin P, Tanner M, Zinsstag J (2003) Brucellosis and Q-fever seroprevalences of nomadic pastoralists and their livestock in Chad. Prev Vet Med 61:279–293

  32. Sciutto E et al (2007) Improvement of the synthetic tri-peptide vaccine (S3Pvac) against porcine Taenia solium cysticercosis in search of a more effective. inexpensive manageable vaccine. Vaccine 25:1368–1378. https://doi.org/10.1016/j.vaccine.2006.10.018

  33. Sesardic D (1993) Synthetic peptide vaccines. J Med Microbiol 39:241–242

  34. Shen Y, Maupetit J, Derreumaux P, Tufféry P (2014) Improved PEP-FOLD approach for peptide and miniprotein structure prediction. J Chem Theor Comput 10:4745–4758

  35. Shibue M, Mant CT, Hodges RS (2005) Effect of anionic ion-pairing reagent concentration (1–60 mM) on reversed-phase liquid chromatography elution behaviour of peptides. J Chromatogr A 1080:58–67

  36. Stevens MG, Tabatabai LB, Olsen SC, Cheville NF (1994) Immune responses to superoxide dismutase and synthetic peptides of superoxide dismutase in cattle vaccinated with Brucella abortus strain 19 or RB51. Vet Microbiol 41:383–389

  37. Strupat K (2005) Molecular weight determination of peptides and proteins by ESI and MALDI. Methods Enzymol 405:1–36

  38. Tabatabai LB, Pugh GW Jr (1994) Modulation of immune responses in Balb/c mice vaccinated with Brucella abortus Cu-Zn superoxide dismutase synthetic peptide vaccine. Vaccine 12:919–924

  39. Trauger SA, Webb W, Siuzdak G (2002) Peptide and protein analysis with mass spectrometry. J Spectrosc 16:15–28

  40. Wang SS (1973) p-alkoxybenzyl alcohol resin and p-alkoxybenzyloxycarbonylhydrazide resin for solid phase synthesis of protected peptide fragments. J Am Chem Soc 95:1328–1333

  41. Wang XJ et al (2010) Preparation of a peptide vaccine against GnRH by a bioprocess system based on asparaginase. Vaccine 28:4984–4988. https://doi.org/10.1016/j.vaccine.2010.05.026

  42. Yu HM, Chen ST, Wang KT (1992) Enhanced coupling efficiency in solid-phase peptide synthesis by microwave irradiation. J Org Chem 57:4781–4784

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This work was supported by Research Fund of the Yildiz Technical University (Project Number: 2014-07-04-YL01) and the TUBITAK MSc Scholarship Program in Priority Areas (2210/C).

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Correspondence to Tayfun Acar.

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Acar, T., Pelit Arayıcı, P., Ucar, B. et al. Synthesis, Characterization and Lipophilicity Study of Brucella abortus’ Immunogenic Peptide Sequence That Can Be Used in the Future Vaccination Studies. Int J Pept Res Ther 25, 911–918 (2019). https://doi.org/10.1007/s10989-018-9739-0

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  • Solid phase peptide synthesis (SPPS)
  • Brucella abortus
  • Peptide vaccine
  • Validation
  • Fluorescence
  • Lipophilicity