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J Allergy Clin Immunol. 1998 May;101(5):691-8.

Superior biologic activity of the recombinant bee venom allergen hyaluronidase expressed in baculovirus-infected insect cells as compared with Escherichia coli.

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1
Zieglerspital and Institute of Biochemistry, University of Bern, Switzerland.

Abstract

BACKGROUND:

Hyaluronidase (Hya) is one of several allergens in honeybee venom. Its cDNA sequence was recently described.

OBJECTIVE:

We sought to express recombinant Hya in prokaryotic and eukaryotic systems and to compare it with natural (n)Hya for biologic activity.

METHODS:

In Escherichia coli Hya was produced as inclusion body 6 x His-fusion protein. In baculovirus-infected insect cells expression was obtained by cotransfection of linearized Bac-N-Blue DNA and pMelBac transfer vector into Spodoptera frugiperda cells.

RESULTS:

Enzymatic activity of Hya from the baculovirus system was equal to nHya, and that of the enzyme expressed in E. coli was only 20% to 30% of nHya. In vitro IgE binding was similar in nHya and the enzyme from baculovirus but markedly lower in Hya expressed in E. coli.

CONCLUSIONS:

Biologic activity of Hya expressed in baculovirus-infected insect cells was comparable with that of the natural enzyme, indicating a native-like conformation of the recombinant protein. In contrast, the enzyme expressed in E. coli as an inclusion-body protein and reconstituted in vitro reached only 20% to 30% of the activity of nHya.

PMID:
9600508
DOI:
10.1016/S0091-6749(98)70179-4
[Indexed for MEDLINE]
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