Allergy Asthma Immunol Res.  2013 Mar;5(2):102-105. 10.4168/aair.2013.5.2.102.

Preparation and Characterization of an Extract of German Cockroach From a Korean Source

Affiliations
  • 1Department of Internal Medicine and Institute of Allergy, Yonsei University College of Medicine, Seoul, Korea. parkjw@yuhs.ac
  • 2Center for Immunology and Pathology, Korea National Institute of Health, Cheongwon, Korea.
  • 3Department of Environmental Medical Biology and Institute of Tropical Medicine, Arthropod of Medical Importance Resource Bank, Yonsei University College of Medicine, Seoul, Korea.

Abstract

PURPOSE
The cockroach (CR) is an important cause of respiratory allergic disorders. We prepared a German CR extract in a standardized way and analyzed its allergenic properties.
METHODS
The extract was prepared from German CR (Blattella germanica) obtained from a Korean colony, and its allergenic activity was compared with that of the commercial Hollister-Stier (HS) extract. The concentrations of Bla g 1 and Bla g 2 were measured, and an in vitro specific IgE binding inhibition assay was performed to assess IgE reactivity. Proteolytic activity was examined by gelatin zymography.
RESULTS
Bla g 1 and Bla g 2 were detected at 405 U/mg and 273 ng/mg, respectively, in the Korean extract, and at 187 U/mg and 56 ng/mg, respectively, in the HS extract. The Korean extract showed 94.2% inhibition of IgE reactivity, as compared with the HS extract. A similar pattern of IgE-reactive bands was detected for the two extracts, indicating that their allergenic components are similar. The proteolytic activities of the Korean and HS extracts were found to be similar in gelatin zymography. The endotoxin levels in the Korean and HS extracts were 3,440 EU/mL and 6,580 EU/mL, respectively.
CONCLUSIONS
The German CR extract was prepared in a standardized way. The extract produced in this study will be useful for the development of allergy diagnostics and immunotherapeutic agents.

Keyword

Allergen; German cockroach; Korea; standardization

MeSH Terms

Allergens
Blattellidae
Cockroaches
Gelatin
Hypersensitivity
Immunoglobulin E
Korea
Allergens
Gelatin
Immunoglobulin E

Figure

  • Fig. 1 Protein analysis of the German cockroach extracts. Allergen extracts (10 µg/well) were separated on a 12% SDS-PAGE gel under reducing conditions (A). The allergenic components of the extracts were compared by IgE immunoblotting (B). Gelatinolytic protease activity was analyzed on a 10% SDS-PAGE gel (0.2 µg/well) that contained 0.1% gelatin (C). M, molecular mass marker; K, Korean German cockroach extract; HS, US German cockroach extract (Hollister-Stier).

  • Fig. 2 Inhibition analysis of the cockroach extracts. The specific IgE antibody binding activities of the German cockroach extracts prepared in Korea (Bg-YS) and the US (Bg-HS) were compared in an in vitro specific IgE antibody binding inhibition assay.


Cited by  1 articles

Allergen standardization
Jung-Won Park, Kyoung Yong Jeong
Allergy Asthma Respir Dis. 2018;6(4):191-196.    doi: 10.4168/aard.2018.6.4.191.


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