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Original Paper

Proteomic Analysis of Putative Latex Allergens

Yagami T.a · Haishima Y.a · Tsuchiya T.a · Tomitaka-Yagami A.b · Kano H.b · Matsunaga K.b

Author affiliations

aDivision of Medical Devices, National Institute of Health Sciences, Tokyo, bDepartment of Dermatology, Fujita Health University School of Medicine, Toyoake, Japan

Related Articles for ""

Int Arch Allergy Immunol 2004;135:3–11

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Article / Publication Details

First-Page Preview
Abstract of Original Paper

Received: December 23, 2003
Accepted: June 07, 2004
Published online: September 22, 2004
Issue release date: September 2004

Number of Print Pages: 9
Number of Figures: 3
Number of Tables: 2

ISSN: 1018-2438 (Print)
eISSN: 1423-0097 (Online)

For additional information: http://www.karger.com/IAA

Abstract

Background: Extensive analysis of allergenic proteins is generally time-consuming and labor-intensive. Accordingly, a rapid and easy procedure for allergen identification is required. As sequence information on proteins and genes is accumulated in databases, it is becoming easier to identify a candidate protein using proteomic strategies, i.e. two-dimensional gel electrophoresis, site-specific fragmentation, mass spectrometry and then database search. In this study, we evaluated the usefulness of a proteomic strategy for identifying putative allergens through its application to latex proteins. Methods: Latex proteins were separated with two-dimensional gel electrophoresis, and putative allergens were visualized by IgE immunoblotting using pooled serum from latex-sensitive patients. The IgE-interactive proteins were cut out from the negatively stained two-dimensional gel and subjected to in-gel digestion by trypsin. Then the resulting peptides were analyzed with mass spectrometry. Based on the mass spectrometric data we obtained, the allergen candidates were assigned by a database search. Results: Five previously reported allergens and five new allergen candidates were identified with the proteomic approach without isolating the individual proteins. Less than 1 mg of crude latex protein was sufficient for the entire protocol. Because plural proteins can be processed in parallel, analysis of about 50 IgE-interactive proteins was accomplished within 1 week. Conclusions: Analysis of putative allergens with proteomic strategies (allergenomics) is a promising avenue for rapid and exhaustive research. The high resolving power of two-dimensional gel electrophoresis is superior to conventional gel electrophoresis. Moreover, the notable sensitivity and speed of mass spectrometry have pronounced advantages over the N-terminal sequencing that has generally been used for protein identification.

© 2004 S. Karger AG, Basel


Article / Publication Details

First-Page Preview
Abstract of Original Paper

Received: December 23, 2003
Accepted: June 07, 2004
Published online: September 22, 2004
Issue release date: September 2004

Number of Print Pages: 9
Number of Figures: 3
Number of Tables: 2

ISSN: 1018-2438 (Print)
eISSN: 1423-0097 (Online)

For additional information: http://www.karger.com/IAA


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