Recombinant production of a peroxidase-protein G fusion protein in Pichia pastoris

Florian Krainer, Barbara Darnhofer, Ruth Birner-Gruenberger, Anton Glieder

Research output: Contribution to journalArticle

Abstract

Streptococcal protein G (SpG) binds immunoglobulin G from a broad range of mammalian species with high affinity. Chemical conjugations of SpG to the reporter enzyme horseradish peroxidase (HRP) are commonly used in immunohistochemical applications. However, commercial HRP preparations are typically isolated from horseradish roots as varying mixtures of HRP isoenzymes with different biochemical properties, and chemical conjugation procedures lead to heterogeneous HRP-SpG preparations, partially including inactivated enzyme. A recombinant process allows the production of a well-defined HRP isoenzyme fused to SpG at constant 1:1 stoichiometry in a single step without the need for laborious chemical conjugation. By using state-of-the-art biotechnological tools, we produced a recombinant HRP-SpG fusion protein in Pichia pastoris in bioreactor cultivations. Purified HRP-SpG was tested successfully for functional binding of antibodies from different mammalian serums. Recombinant production of this novel well-defined fusion protein follows quality-by-design principles and facilitates the production of more reliable and cost-effective diagnostic kits.

LanguageEnglish
Pages24-27
Number of pages4
JournalJournal of biotechnology
Volume219
DOIs
StatusPublished - 10 Feb 2016

Fingerprint

Pichia
Horseradish Peroxidase
Peroxidase
Proteins
Isoenzymes
Armoracia
Bioreactors
Enzymes
Streptococcus IgG Fc-binding protein
Immunoglobulin G
Costs and Cost Analysis
Antibodies
Serum

Keywords

  • Antibody
  • Diagnostics
  • Fusion protein
  • Peroxidase
  • Protein G

ASJC Scopus subject areas

  • Biotechnology
  • Applied Microbiology and Biotechnology

Cite this

Recombinant production of a peroxidase-protein G fusion protein in Pichia pastoris. / Krainer, Florian; Darnhofer, Barbara; Birner-Gruenberger, Ruth; Glieder, Anton.

In: Journal of biotechnology, Vol. 219, 10.02.2016, p. 24-27.

Research output: Contribution to journalArticle

Krainer, Florian ; Darnhofer, Barbara ; Birner-Gruenberger, Ruth ; Glieder, Anton. / Recombinant production of a peroxidase-protein G fusion protein in Pichia pastoris. In: Journal of biotechnology. 2016 ; Vol. 219. pp. 24-27
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