Application of western blot analysis for detection of prolamin proteins in cereal grains and bread

Authors

  • Peter Socha Department of Biochemistry and Biotechnology, Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2, 949 76 Nitra
  • Barbara Mickowska Malopolska Centre of Food Monitoring and Certification, Faculty of Food Technology, Agriculture University in Cracow, Balicka 122, 30-149 Cracow
  • Elżbieta Mazur Malopolska Centre of Food Monitoring and Certification, Faculty of Food Technology, Agriculture University in Cracow, Balicka 122, 30-149 Cracow
  • Dana Urminská Department of Biochemistry and Biotechnology, Faculty of Biotechnology and Food Sciences, Slovak University of Agriculture in Nitra, Tr. A. Hlinku 2, 949 76 Nitra
  • Ewa Cieślik Malopolska Centre of Food Monitoring and Certification, Faculty of Food Technology, Agriculture University in Cracow, Balicka 122, 30-149 Cracow

DOI:

https://doi.org/10.5219/115

Keywords:

Western blot, prolamins, cereal, celiac disease

Abstract

Celiac disease is an inflammatory condition of the small intestine in genetically susceptible individuals caused by ingestion of wheat gluten and corresponding proteins from barley and rye. Cereal storage proteins (prolamins) are responsible for immunological response of patients with celiac disease. Prolamins are alcohol soluble fractions, namely gliadins (wheat), hordeins (barley) and secalins (rye). The main triggering factor is wheat fraction with low molecular weight (20-30 kDa) called α-gliadins. Immunochemical detection of celiac active proteins is based on reactivity of gluten-detecting antibodies with prolamins extracted from cereals. In our study, we used Western blot analysis for detection of prolamin complex in cereal grains and processed foods (breads). Western blot was carried out by polyclonal antibody raised against wheat gluten. Reaction was positive for all kind of cereal grains. The samples of wheat and spelt wheat show much more positive affinity to antibody than rye and oat. As well as for cereal grains, all samples of bread showed positive immunological reaction with used antibody. Western blot analysis with gluten polyclonal antibody is suitable method for qualitative detection of prolamin complex in cereal grains and processed foods.

Downloads

Download data is not yet available.

References

ARENDT, E. K., DAL BELLO, F., 2008. Gluten-free cereal products and beveragep. USA: Elsevier Inc., 2008, 445 p. ISBN 978-0-12-373739-7.

ARENTZ-HANSEN, H., FLECKENSTEIN, B., MOLBERG, Ø., SCOTT, H., KONING, F., JUNG, G., ROEPSTORFF, P., LUNDIN, K. E. A., SOLLID, L. M., 2004. The molecular basis for oat intolerance in patients with celiac disease. In PLoS Medicine, vol. 1, 2004, no. 1, p. 84-92.

PMid: 15526039

BATTAIS, F., PINEAU, F., POPINEAU, Y., APARICIO, C., KANNY, G., GUERIN, L., MONERET-VAUTRIN, D. A., DENERY-PAPINI, P., 2003. Food allergy to wheat: identification of immunoglobulin E and immunoglobulin G-binding proteins with sequential extracts and purified proteins from wheat flour. In Clinical &Experimental Allergy, vol. 33, 2003, no. 7, p. 962-970.

PMid: 12859454

CIACCI, C., CIRILLO, M., CAVALLARO, R., MAZZACCA, G., 2002. Long-term follow-up of celiac adults on gluten-free diet: prevalence and correlates of intestinal damage. In Digestion, vol. 66, 2002, no. 3, p. 178-185.

PMid: 12481164

CICCOCIOPPO, R., DI SABATINO, A., CORAZZA, G. R., 2005. The immune recognition of gluten in celiac disease. In Clinical and Experimental Immunology, vol. 140, 2005, no. 3, p. 408-416.

PMid: 15932501

EBO, D. G., STEVENS, W. J., 2001. IgE-mediated food allergy – extensive review of the literature. InActa Clinica Belgica, vol. 56, 2001, no. 4, p. 234-247.

PMid: 11603253

ECKERT van, R., BOND, J., RAWSON, P., KLEIN, CH. L., STERN, M., JORDAN, T. W., 2010. Reactivity of gluten detecting monoclonal antibodies to a gliadin reference material. In Journal of Cereal Science, vol. 51, 2010, no. 2, p. 198-204.

https://doi.org/10.1016/j.jcs.2009.11.012

ELLIS, H. J., ROSEN-BRONSON, P., O’REILLY, N., CICLITIRA, P. J., 1998. Measurement of gluten using a monoclonal antibody against a coeliac toxic peptide of A gliadin. In Gut, vol. 43, 1998, no. 2, p. 190-195.

HILL, P. G., McMILLAN, P. A., 2006. Anti-tissue transglutaminase antibodies and their role in the investigation of coeliac disease. In Annals of Clinical Biochemistry, vol. 43, 2006, no. 2, p. 105-117.

PMid: 16536912

JANATUINEN, E. K., PIKKARAINEN, P. H., KEMPPAINEN, T. A., KOSMA, V. M., JÄRVINEN, R. M. K., UUSITUPA, M. I. J., JULKUNEN, R. J. K., 1995. A comparison of diets with and without oats in adults with celiac disease. In The New England Journal of Medicine, vol. 333, 1995, no. 16, p. 1033-1037.

PMid: 7675045

JANATUINEN, E. K., KEMPPAINEN, T. A., JULKUNEN, R. J. K., KOSMA, V. M., MÄKI, M., HEIKKINEN, M., UUSITUPA, M. I. J., 2002. No harm from five year ingestion of oats in coeliac disease. In Gut, vol. 50, 2002, no. 3, p. 332-335.

PMid: 11839710

LUNDIN, K. E. A., NILSEN, E. M., SCOTT, H. G., LØBERG, E. M., GJØEN, A., BRATLIE, J., SKAR, V., MENDEZ, E., LØVIK, A., KETT, K., 2003. Oats induced villous atrophy in coeliac disease. In Gut, vol. 52, 2003, no. 11, p. 1649-1652.

PMid: 14570737

PETR, J., MICHALÍK, I., TLASKALOVÁ, H., CAPOUCHOVÁ, I., FAMĚRA, O., URMINSKÁ, D., TUČKOVÁ, L., KNOBLOCHOVÁ, H., 2003. Extention of the spectra of plant products for the diet in coeliac disease. In Czech Journal of Food Sciences, vol. 21, 2003, no. 2, p. 59-70.

PRUSKA-KĘDZIOR, A., KĘDZIOR, Z., GORĄCY, M., PIETROWSKA, K., PRZYBYLSKA, A., SPYCHALSKA, K., 2008. Comparison of rheological, fermentative and baking properties of gluten-free dough formulations. In European Food Research and Technology, vol. 227, 2008, no. 5, p. 1523-1536.

SCIARINI, L. P., RIBOTTA, P. D., LEÓN, A. E., PÉREZ, G. T., 2010. Influence of gluten-free flours and their mixtures on batter properties and bread quality. InFood and Bioprocess Technology, vol. 3, 2010, no. 4, p. 577-585.

SHEWRY, P. R., HALFORD, N. G., 2002. Cereal seed storage proteins: structure, properties and role in grain utilization. In Journal of Experimental Botany, vol. 53, 2002, no. 370, p. 947-958.

https://doi.org/10.1093/jexbot/53.370.947

SCHÄGGER, H., VON JAGOW, G., 1987. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. In Analytical Biochemistry, vol. 166, 1987, no. 2, p. 368-379.

PMid: 2449095

SILANO, M., BENEDETTO, R., MAIALETTI, F., VINCENZI, A., CALCATERRA, R., CORNELL, H. J., VINCENZI, M., 2007. Avenins from different cultivars of oats elicit response by coeliac peripheral lymphocytes. In Scandinavian Journal of Gastroenterology, vol. 42, 2007, no. 11, p. 1302-1305.

PMid: 17852883

STAROVIČOVÁ, M., GÁLOVÁ, Z., KNOBLOCHOVÁ, H., 2003. Identification of glutenin marker in cultivars of three wheat species. In Czech Journal of Genetics and Plant Breeding, vol. 39, 2003, no. 2, p. 51-57.

STØRSRUD, P., HULTHÉN, L. R., LENNER, R. A., 2003. Beneficial effects of oats in the gluten-free diet of adults with special reference to nutrient status, symptoms and subjective experiences. In British Journal of Nutrition, vol. 90, 2003, no. 1, p. 101-107.

PMid: 12844381

URMINSKÁ, D., SOCHA, P., VOLLMANNOVÁ, A., 2009. ELISA and PAGE analysis of protein determinants from cereal and pseudocereal grain causing human coeliac disease. In FEBS Journal, vol. 276, 2009, no. 1, p. 94.

VACCINO, P., BECKER, H. A., BRANDOLINI, A., SALAMINI, F., KILIAN, B., 2009. A catalogue ofTriticum monococcum genes encoding toxic and immunogenic peptides for celiac disease patients. InMolecular Genetics and Genomics, vol. 281, 2009, no. 3, p. 289-300.

PMCid: PMC2757618

WAGA, J., WĘGRZYN, P., 2000. Relationship between some gliadin protein subunits and variation of agronomic traits winter wheat cultivars and strains. In Biuletyn IHAR, vol. 215, 2000, p. 61-67.

WAGA, J., ZIENTARSKI, J., 2007. Isolation and purification of individual gliadin proteins by preparative acid polyacrylamide gel electrophoresis (A-PAGE) for allergenic research. In Polish Journal of Food and Nutrition Sciences, vol. 57, 2007, no. 1, p. 91-96.

WEBER, D., CLÉROUX, CH., GODEFROY, P. B., 2009. Emerging analytical methods to determine gluten markers in processed foods – method development in support of standard setting. In Analytical and Bioanalytical Chemistry, vol. 395, 2009, no. 1, p. 111-117.

PMid: 19636545

WIESER, H., 2007. Chemistry of gluten proteins. InFood Microbiology, vol. 24, 2007, no. 2, p. 115-119.

https://doi.org/10.1016/j.fm.2006.07.004

WEISER, H., KOEHLER, P., 2008. The biochemical basis of celiac disease. In Cereal Chemistry, vol. 85, 2008, no. 1, p. 1-14.

XIE, Z., WANG, C., WANG, K., WANG, P., LI, X., ZHANG, Z., MA, W., YAN, Y., 2010. Molecular characterization of the celiac disease epitope domains in a-gliadin genes in Aegilops tauschii and hexaploid wheats (Triticum aestivum L.). InTheoretical and Applied Genetics, vol. 121, 2010, no. 7, p. 1239-1251.

PMid: 20556595

YALÇIN, E., 2010. Effect of partial removal of prolamins on some chemical and functional properties of barley flours. InFood Science and Biotechnology, vol. 19, 2010, no. 3, p. 735-742.

https://doi.org/10.1007/s10068-010-0103-1

ZINGONE, F., CAPONE, P., CIACCI, C., 2010. Celiac disease: Alternatives to a gluten free diet. In World Journal of Gastrointestinal Pharmacology and Therapeutics, vol. 1, 2010, no. 1, p. 36-39.

https://doi.org/10.4292/wjgpt.v1vi1.36

Potravinárstvo 1/2011

Downloads

Published

2011-01-16

How to Cite

Socha, P. ., Mickowska, B. ., Mazur, E. ., Urminská, D. ., & Cieślik, E. . (2011). Application of western blot analysis for detection of prolamin proteins in cereal grains and bread. Potravinarstvo Slovak Journal of Food Sciences, 5(1), 51-55. https://doi.org/10.5219/115

Issue

Vol. 5 No. 1 (2011): Potravinarstvo

Section

Articles

License

Authors who publish with this journal agree to the following terms:

  1. Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.

  2. Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.

  3. Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).