1.Optimization of Allergen Standardization.
Kyoung Yong JEONG ; Chein Soo HONG ; Joo Shil LEE ; Jung Won PARK
Yonsei Medical Journal 2011;52(3):393-400
Preparation of high quality allergen extracts is essential for the diagnosis and immunotherapy of allergic disorders. Standardization of allergen extracts concerns determination of the allergen unit, development of reference material and measurement of the overall IgE binding capacity of an allergen extract. Recently, quantification of individual allergens has been the main focus of allergen standardization because the allergenicity of most allergen extracts is known to be mainly dependent on the content of a small number of allergen molecules. Therefore, characterization of major allergens will facilitate the standardization of allergens. In this article, we review the current state of allergen standardization. In addition, we briefly summarize the components of allergen extracts that should be under control for the optimization of allergen standardization, since its adjuvant-like activities could play an important role in allergic reactions even though the molecule itself does not bind to the IgE antibodies from subjects.
Adjuvants, Immunologic/chemistry
;
Allergens/chemistry/*immunology/isolation & purification
;
Polymorphism, Genetic
;
Reference Standards
;
Republic of Korea
2.Identification of the major allergen of Macrobrachium rosenbergii (giant freshwater prawn).
Zailatul Hani Mohamad YADZIR ; Rosmilah MISNAN ; Noormalin ABDULLAH ; Faizal BAKHTIAR ; Masita ARIP ; Shahnaz MURAD
Asian Pacific Journal of Tropical Biomedicine 2012;2(1):50-54
OBJECTIVETo characterize the major allergens of Macrobrachium rosenbergii (giant freshwater prawn).
METHODSRaw and cooked extracts of the giant freshwater prawn were prepared. The IgE reactivity pattern was identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting technique with the sera of 20 skin prick test (SPT) positive patients. The major allergen identified was then characterized using the proteomics approach involving a combination of two-dimensional (2-DE) electrophoresis, mass spectrometry and bioinformatics tools.
RESULTSSDS-PAGE of the raw extract showed 23 protein bands (15-250 kDa) but those ranging from 40 to 100 kDa were not found in the cooked extract. From immunoblotting experiments, raw and cooked extracts demonstrated 11 and 5 IgE-binding proteins, respectively, with a molecular mass ranging from 15 to 155 kDa. A heat-resistant 36 kDa protein was identified as the major allergen of both extracts. In addition, a 42 kDa heat-sensitive protein was shown to be a major allergen of the raw extract. The 2-DE gel fractionated the prawn proteins to more than 50 different protein spots. Of these, 10 spots showed specific IgE reactivity with patients' sera. Matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) analysis led to identification of 2 important allergens, tropomyosin and arginine kinase.
CONCLUSIONSIt can be concluded that the availability of such allergens would help in component-based diagnosis and therapy of prawn allergies.
Allergens ; isolation & purification ; Animals ; Computational Biology ; Electrophoresis, Gel, Two-Dimensional ; Electrophoresis, Polyacrylamide Gel ; Fresh Water ; Humans ; Immunoblotting ; Immunoglobulin E ; immunology ; Mass Spectrometry ; Molecular Weight ; Palaemonidae ; chemistry