Production and Characterization of Monoclonal Antibody against Vit v1: A Grape Allergen Belonging to Lipid Transfer Protein Family

  • Mitra Hosseinpour 15 Khordad Hospital, ShahidBeheshti University of Medical Sciences, Tehran, Iran AND Immuno-Biochemistry Lab, Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran
  • Kazem Mashayekhi Immuno-Biochemistry Lab, Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran
  • Reza Falak Department of Immunology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran
  • Sirous Jamalzehi Department of Medical Laboratory Sciences, Iranshahr University of Medical Sciences, Iranshahr, Iran
  • Saeed Mohammadian Haftcheshmeh Department of Medical Immunology, Nanotechnology Research Center, Bu-Ali Research Institute, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran
  • Mohamad Javad Mousavi Bushehr University of Medical Sciences, Bushehr, Iran AND Department of Immunology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
  • Anvar Soleimani Department of Clinical Biochemistry, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran
  • Khadijeh Koushki Immuno-Biochemistry Lab, Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran
  • Mojtaba Sankian Mail Immuno-Biochemistry Lab, Immunology Research Center, Mashhad University of Medical Sciences, Mashhad, Iran
  • Mohammad Soukhtanloo Department of Clinical Biochemistry, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran
Keywords:
Allergy, Grape, Lipid transfer protein, Monoclonal antibody, Non-specific lipid transfer protein

Abstract

Allergy to non-specific lipidtransfer protein (nsLTP), the major allergen of grape (Vit v1), is considered as one of the most common fruit allergies in Iran. Therefore, a specific monoclonal antibody (mAb) can be used for the characterization and assessment of. Accordingly, this study aimed to generate and characterize a mAb against Vit v1 with a diagnostic purpose. To this end, Vit v1 allergen (9 kDa) was extracted using a modified Bjorksten extraction method. Natural Vit v1-immunized mouse splenocytes were fused with SP2/0Ag-14 myeloma cells for generating hybridoma cells. Specific antibody-secreting Hybridoma cells were selected using ELISA. Finally, anti-Vit v1 mAb was characterized by western blotting, ELISA, and isotyping methods. In the current study, a 9 kDa (Vit v1) protein was attained fromcrude and fresh juice of grape extracts and the isotype of desired anti-Vit v1 mAb was determined as IgM with k light chain. In addition, The ELISA results demonstrated that anti-Vit v1 mAb was specified against natural Vit v1 in the grape cultivar and related LTP allergens, such as Pla or 3 (p<0.0001). In the present study, a specific mAb was produced for detecting the LTP allergen. This mAb with a confirmed specificity can be utilized for evaluating the LTP allergens and their allergenicity in different grape cultivars.

References

1. Vieths S, Scheurer S, Ballimer-Weber B. Current understanding of cross‐reactivity of food allergens and pollen. Ann N Y Acad Sci. 2002;964(1):47-68.
2. Takamatsu N, Kondo Y, Tsuge I, Nakajima Y, Naruse N, Tanaka K, et al. A study of cross-reactivity between citrus fruit and pollen allergens in oral allergy syndrome and food-dependent exercise-induced anaphylaxis in Japan. Fujita Medical Journal. 2016;2(1):6-11.
3. Zuidmeer L, Goldhahn K, Rona RJ, Gislason D, Madsen C, Summers C, et al. The prevalence of plant food allergies: a systematic review. J Allergy Clin Immunol. 2008;121(5):1210-8. e4.
4. Nwaru BI, Hickstein L, Panesar S, Muraro A, Werfel T, Cardona V, et al. The epidemiology of food allergy in Europe: a systematic review and meta‐analysis. Allergy. 2014;69(1):62-75.
5. Sankian M, Bagheri Y, Vahedi F, Azad FJ, Varasteh A-R. Validity of using recombinant melon profilin, Cuc m 2, for diagnosis of melon allergy. Rep Biochem Mol Biol. 2012;1(1):14.
6. Sankian M, Mahmoudi M, Varasteh A-R. Molecular cloning, characterization, and expression of Cuc m 2, a major allergen in Cucumis melo. Rep Biochem Mol Biol. 2013;1(2):49.
7. Falak R, Sankian M, Tehrani M, Azad FJ, Abolhasani A, Varasteh A. Clinical and laboratory investigation of oral allergy syndrome to grape. Iran J Allergy Asthma Immunol. 2012;11(2):147-55.
8. Falak R, Sankian M, Noorbakhsh R, Tehrani M, Assarehzadegan MA, Jabbari Azad F, et al. Identification and characterisation of main allergic proteins in Vitis vinifera vitis. Food Agr Immunol. 2013;24(3):255-68.
9. Kalogeromitros DC MM, Gregoriou SG, Katoulis AC, Straurianeas NG. Sensitization to other foods in subjects with reported allergy to grapes. Allergy Asthma Proc. 2006;27(1).
10. Karakaya G, Kalyoncu A. Allergy to grapes. Ann Allergy Asthma Immunol. 2000;84(2):265.
11. Asero R, Mistrello G, Roncarolo D, de Vries SC, Gautier M-Fo, Ciurana CL, et al. Lipid transfer protein: a pan-allergen in plant-derived foods that is highly resistant to pepsin digestion. Int Arch Allergy Immunol. 2000;122(1):20-32.
12. Finkina E, Melnikova D, Bogdanov I. Lipid transfer proteins as components of the plant innate immune system: structure, functions, and applications. Acta Naturae. 2016;8(2 (29)).
13. Van Loon L, Van Strien E. The families of pathogenesis-related proteins, their activities, and comparative analysis of PR-1 type proteins. Physiol Mol Plant P. 1999;55(2):85-97.
14. I Finkina E, N Melnikova D, V Bogdanov I, V Ovchinnikova T. Plant pathogenesis-related proteins PR-10 and PR-14 as components of innate immunity system and ubiquitous allergens. Curr Med Chem. 2017;24(17):1772-87.
15. Pastorello EA, Farioli L, Pravettoni V, Ortolani C, Fortunato D, Giuffrida MG, et al. Identification of grape and wine allergens as an endochitinase 4, a lipid-transfer protein, and a thaumatin. J Allergy Clin Immunol. 2003;111(2):350-9.
16. Mari A, Rasi C, Palazzo P, Scala E. Allergen databases: current status and perspectives. Curr Allergy Asthma Rep. 2009;9(5):376-83.
17. Jaeckels N, Bellinghausen I, Fronk P, Heydenreich B, Saloga J, Decker H. Assessment of sensitization to grape and wine allergens as possible causes of adverse reactions to wine: a pilot study. Clin Transl Allergy. 2015;5(1):21.
18. Akkerdaas JH, Wensing M, Knulst AC, Krebitz M, Breiteneder H, de Vries S, et al. How accurate and safe is the diagnosis of hazelnut allergy by means of commercial skin prick test reagents? Int Arch Allergy Immunol. 2003;132(2):132-40.
19. Brenna O, Pompei C, Ortolani C, Pravettoni V, Pastorello EA, Farioli L. Technological processes to decrease the allergenicity of peach juice and nectar. J Agr Food Chem. 2000;48(2):493-7.
20. Chapman MD. Allergen specific monoclonal antibodies: new tools for the management of allergic disease. Allergy. 1988;43:7-14.
21. Björkstén F, Halmepuro L, Hannuksela M, Lahti A. Extraction and properties of apple allergens. Allergy. 1980;35(8):671-7.
22. Soukhtanloo M, Falak R, Sankian M, Varasteh A-R. Generation and characterization of anti-chitinase monoclonal antibodies. Hybridoma. 2011;30(2):145-51.
23. Pazouki N, Sankian M, Leung PT, Nejadsattari T, Khavari-Nejad R-A, Varasteh A-R. Identification of cyclophilin as a novel allergen from Platanus orientalis pollens by mass spectrometry. J Biosci Bioeng. 2009;107(2):215-7.
24. Varasteh A, Pazouki N, Sankian M. Cloning, Expression, and Immunological Characterization of Recombinant Platanus orientalis Allergen Pla or 3.0101. J Allergy Clin Immunol. 2009;123(2):S228.
25. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. nature. 1970;227(5259):680-5.
26. Nelson MR CL. Allergen immunotherapy extract preparation manual. AAAAI Practice Management Resource Guide. 2012:1-39.
27. Sánchez-Monge R, Lombardero M, García-Sellés FJ, Barber D, Salcedo G. Lipid-transfer proteins are relevant allergens in fruit allergy. J Allergy Clin Immunol. 1999;103(3):514-9.
28. Monteiro S, Piçarra-Pereira M, Mesquita P, Loureiro V, Teixeira A, Ferreira R. The wide diversity of structurally similar wine proteins. J Agr Food Chem. 2001;49(8):3999-4010.
29. Vincent D, Wheatley MD, Cramer GR. Optimization of protein extraction and solubilization for mature grape berry clusters. Electrophoresis. 2006;27(9):1853-65.
30. Jona R, Fronda A. Comparative histochemical analysis of cell wall polysaccharides by enzymatic and chemical extractions of two fruits. Biotech Histochem. 1997;72(1):22-8.
31. Vassilopoulou EV, Zuidmeer L, Akkerdaas J, Rigby N, Moreno FJ, Papadopoulos NG, et al. Optimized techniques for the extraction of grape allergens appropriate for in vivo and in vitro testing and diagnosis. Mo. Nutr Food Res. 2007;51(3):360-6.
32. Solá RJ, Griebenow K. Effects of glycosylation on the stability of protein pharmaceuticals. J Pharm Sci. 2009;98(4):1223-45.
33. García B, Lizaso M. 1 Cross-reactivity Syndromes in Food Allergy. J Investig Allergol Clin Immunol. 2011;21(3):162.
34. Gao Z-s, Ma Y-t, Zhou X, Yang Z-w, Jia H-j, Gao L, et al. Quantification of peach fruit allergen lipid transfer protein by a double monoclonal antibody-based sandwich ELISA.Food Anal Methods. 2016;9(4):823-30.
35. Murakami-Yamaguchi Y, Hirose J, Kizu K, Okazaki F, Fujii W, Narita H. Quality control system for beer developed with monoclonal antibodies specific to barley lipid transfer protein. Antibodies. 2012;1(3):259-72.
36. Ritter MA. Polyclonal and monoclonal antibodies. Diagnostic and Therapeutic Antibodies. 2000:23-34.
37. Sankian M, Glosaze Shirazi F, Arafi M, Moghadam M, Varasteh A. Production and characterization of monoclonal antibody against Saffron pollen profilin, Cro s 2.Iran J Immunol. 2008;5(3):156-62.
38. Jaeckels N, Tenzer S, Rosfa S, Schild H, Decker H, Wigand P. Purification and structural characterisation of lipid transfer protein from red wine and grapes. Food Chem. 2013;138(1):263-9.
39. Farshidi N, Moghaddam M, Yaghoubi V, Ayati SH, Varasteh A-r, Sankian M. Type IV chitinase quantification in four different grape cultivars (Vitis vinifera) in northeast of Iran by an indirect sandwich enzyme-linked immunosorbent assay. J Immunoass Immunoch. 2019;40(2):139-48.
Published
2020-04-16
How to Cite
1.
Hosseinpour M, Mashayekhi K, Falak R, Jamalzehi S, Mohammadian Haftcheshmeh S, Mousavi MJ, Soleimani A, Koushki K, Sankian M, Soukhtanloo M. Production and Characterization of Monoclonal Antibody against Vit v1: A Grape Allergen Belonging to Lipid Transfer Protein Family. Iran J Allergy Asthma Immunol. 19(2):139-148.
Section
Original Article(s)