Expression of tropomyosin from Blattella germanica as a recombinant non-fusion protein in Pichia pastoris and comparison of its IgE reactivity with its native counterpart

Kyoung Yong Jeong, Jongweon Lee, In Yong Lee, Chein Soo Hong, Han Il Ree, Tai Soon Yong

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Tropomyosins derived from invertebrates are well-known pan allergens. However, the allergenicities of recombinant tropomyosins are variable. Here, we undertook to compare the IgE-binding reactivities of native and recombinant German cockroach tropomyosins. Native tropomyosin was purified by ammonium sulfate fractionation, hydroxyapatite column chromatography, and electroelution, and recombinant tropomyosin was expressed in Pichia pastoris. The allergenicities of the native and recombinant tropomyosins were compared by ELISA inhibition analysis. Native German cockroach tropomyosin showed 18% IgE-binding reactivity to German cockroach sensitized sera. Recombinant tropomyosin was produced without fusion protein and its N-terminus was blocked like that of the native counterpart. The IgE-binding reactivity of the recombinant was found to be comparable to that of native tropomyosin over the concentration range 1-1000ng/ml by ELISA inhibition testing. Recombinant German cockroach tropomyosin expressed in Pichia pastoris showed better allergenicity than that expressed in Escherichia coli. Other factors in addition to the structural differences of native and recombinant proteins may also influence the IgE reactivities of tropomyosins.

Original languageEnglish
Pages (from-to)273-278
Number of pages6
JournalProtein Expression and Purification
Volume37
Issue number2
DOIs
Publication statusPublished - 2004 Oct 1

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Recombinant Fusion Proteins
Tropomyosin
Pichia
Immunoglobulin E
Blattellidae
Enzyme-Linked Immunosorbent Assay
Ammonium Sulfate
Invertebrates
Durapatite
Recombinant Proteins
Allergens
Chromatography

All Science Journal Classification (ASJC) codes

  • Biotechnology

Cite this

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title = "Expression of tropomyosin from Blattella germanica as a recombinant non-fusion protein in Pichia pastoris and comparison of its IgE reactivity with its native counterpart",
abstract = "Tropomyosins derived from invertebrates are well-known pan allergens. However, the allergenicities of recombinant tropomyosins are variable. Here, we undertook to compare the IgE-binding reactivities of native and recombinant German cockroach tropomyosins. Native tropomyosin was purified by ammonium sulfate fractionation, hydroxyapatite column chromatography, and electroelution, and recombinant tropomyosin was expressed in Pichia pastoris. The allergenicities of the native and recombinant tropomyosins were compared by ELISA inhibition analysis. Native German cockroach tropomyosin showed 18{\%} IgE-binding reactivity to German cockroach sensitized sera. Recombinant tropomyosin was produced without fusion protein and its N-terminus was blocked like that of the native counterpart. The IgE-binding reactivity of the recombinant was found to be comparable to that of native tropomyosin over the concentration range 1-1000ng/ml by ELISA inhibition testing. Recombinant German cockroach tropomyosin expressed in Pichia pastoris showed better allergenicity than that expressed in Escherichia coli. Other factors in addition to the structural differences of native and recombinant proteins may also influence the IgE reactivities of tropomyosins.",
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Expression of tropomyosin from Blattella germanica as a recombinant non-fusion protein in Pichia pastoris and comparison of its IgE reactivity with its native counterpart. / Jeong, Kyoung Yong; Lee, Jongweon; Lee, In Yong; Hong, Chein Soo; Ree, Han Il; Yong, Tai Soon.

In: Protein Expression and Purification, Vol. 37, No. 2, 01.10.2004, p. 273-278.

Research output: Contribution to journalArticle

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