High-level secretory expression of immunologically active intact antibody from the yeast Pichia pastoris

Abiodun A. Ogunjimi, John M. Chandler, Christopher M. Gooding, Adrian Recinos, Prabhakara V Choudary

Research output: Contribution to journalArticle

26 Scopus citations

Abstract

We have produced a functional murine antibody to dioxin in the culture medium of the methylotrophic yeast Pichia pastoris. Complementary DNA copies encoding the light (κ) and heavy (γ) chains of the dioxin monoclonal antibody, DD1, were each placed under the control of P. pastoris alcohol oxidase (AOXI) promoter and Saccharomyces cerevisiae α-mating factor secretion signal sequence. The resulting expression cassettes were assembled into a single plasmid (pPICZαDD1) to permit co-expression of both light and heavy chains of the antibody molecule. P. pastor is SMD1168 (pep4, his4) transformed with pPICZαDD1 was able to secrete intact antibody into the culture medium. As high as 36 mg l-1 of the antibody was produced in shake-flask cultures after 96-h induction with methanol. Functional analysis using immunoassay confirmed murine nature of the recombinant antibody and its ability to bind dioxin.

Original languageEnglish (US)
Pages (from-to)561-567
Number of pages7
JournalBiotechnology Letters
Volume21
Issue number6
DOIs
StatePublished - 1999

Keywords

  • Antibody
  • Dioxin monoclonal antibody
  • Expression
  • Pichia pastoris
  • Yeast

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology
  • Microbiology

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