Next Article in Journal
Acknowledgement to Reviewers of Foods in 2014
Previous Article in Journal
Effect of Food and Vitamin D Supplements on the Serum 25(OH)D3 Concentration in Children during Winter Months
Article Menu

Export Article

Open AccessArticle
Foods 2014, 3(4), 642-657; doi:10.3390/foods3040642

Purification of Recombinant Peanut Allergen Ara h 1 and Comparison of IgE Binding to the Natural Protein

Southern Regional Research Center, Agricultural Research Service, United States Department of Agriculture, 1100 Robert E. Lee, Blvd., New Orleans, LA 70124, USA
Current address: Department of Biology, University of New Orleans, New Orleans, LA 70148, USA.
Current address: Department of Medicine, Louisiana State University Health Sciences Center, New Orleans, LA 70112, USA.
*
Author to whom correspondence should be addressed.
Received: 23 October 2014 / Revised: 24 November 2014 / Accepted: 29 November 2014 / Published: 18 December 2014
View Full-Text   |   Download PDF [610 KB, uploaded 18 December 2014]   |  

Abstract

Allergic reactions to food are on the rise worldwide and there is a corresponding increase in interest to understand the molecular mechanisms responsible. Peanut allergies are the most problematic because the reaction often persists into adulthood and can be as severe as anaphylaxis and death. The purpose of the work presented here was to develop a reproducible method to produce large quantities of pure recombinant Ara h 1(rAra h 1) that will enable standardization of immunological tests for patients and allow structural and immunological studies on the wild type and mutagenized forms of the protein. Ara h 1 is initially a pre-pro-protein which, following two endoproteolytic cleavages, becomes the mature form found in peanut. The mature form however has flexible regions that make it refractory to some structural studies including crystallography. Therefore, independent purification of the mature and core regions was desirable. Expression constructs were synthesized cDNA clones for each in a pET plasmid vector without tags. Codons were optimized for expression in E. coli. High-level expression was achieved in BL21 strains. Purification to near homogeneity was achieved by a combination of ammonium sulfate precipitation and ion exchange chromatography. The purified rAra h 1 was then compared with natural Ara h 1 for IgE binding. All patients recognized both the folded natural and rAra h 1, but the IgE binding to the rArah1 was significantly reduced in comparison to the natural allergen, which could potentially make it useful for immunotherapeutic purposes. View Full-Text
Keywords: peanut allergy; immunodominant; Ara h 1; IgE binding peanut allergy; immunodominant; Ara h 1; IgE binding
Figures

Figure 1

This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

Scifeed alert for new publications

Never miss any articles matching your research from any publisher
  • Get alerts for new papers matching your research
  • Find out the new papers from selected authors
  • Updated daily for 49'000+ journals and 6000+ publishers
  • Define your Scifeed now

SciFeed Share & Cite This Article

MDPI and ACS Style

Hurlburt, B.K.; McBride, J.K.; Nesbit, J.B.; Ruan, S.; Maleki, S.J. Purification of Recombinant Peanut Allergen Ara h 1 and Comparison of IgE Binding to the Natural Protein. Foods 2014, 3, 642-657.

Show more citation formats Show less citations formats

Related Articles

Article Metrics

Article Access Statistics

1

Comments

[Return to top]
Foods EISSN 2304-8158 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top