High-level secretion of two antibody single chain Fv fragments by Pichia pastoris

Patrick Eldin, Mary E. Pauza, Yoko Hieda, Gaofeng Lin, Michael P. Murtaugh, Paul R. Pentel, Christopher A. Pennell

Research output: Contribution to journalArticlepeer-review

73 Scopus citations

Abstract

The diagnostic and therapeutic applications of antibody single-chain Fv (sFv) fragments often require large amounts of protein that can be problematic and expensive to obtain. Here we report the secretion of two sFv fragments by the yeast Pichia pastoris at levels up to 250 mg/l. Soluble sFv fragments were purified from culture supernatants in one step by affinity or metal-chelating chromatography, and were indistinguishable from their bacterially expressed counterparts in terms of affinity. Secretion of functional sFv fragments by Pichia pastoris provides a low cost, high yield alternative to current sFv expression systems.

Original languageEnglish (US)
Pages (from-to)67-75
Number of pages9
JournalJournal of Immunological Methods
Volume201
Issue number1
DOIs
StatePublished - Feb 14 1997

Bibliographical note

Funding Information:
We thank Drs. Barton Haynes (Duke University), Tucker LeBien (University of Minnesota), and Ray Mernaugh (Pharmacia Biotech) for kindly providing the 3A1f hybridoma, the biotinylated antibodies, and the anti-E tag affinity column, respectively. G.L. was supported by a National Institute of General Medical Sciences Biotechnology Training Fellowship (IT32-GM08347). P.E. was sponsored by a postdoctoral fellowship from Elf Aquitaine, Inc. (France) while M.E.P. was supported by a predoctoral fellowship from an NIH Immunology Training Grant T3 AI-07313 and by a Doctoral Dissertation Fellowship from the University of Minnesota. This work was supported in part by grants to C.A.P. from the National Cancer Institute (CA-59510), the Leukemia Task Force, and the American Cancer Society, and by grant from the National Institute of Mental Health (MH-42799) to P.R.P.

Copyright:
Copyright 2007 Elsevier B.V., All rights reserved.

Keywords

  • CD7
  • Pichia pastoris
  • desipramine
  • single chain Fv
  • surface plasmon resonance

Fingerprint Dive into the research topics of 'High-level secretion of two antibody single chain Fv fragments by Pichia pastoris'. Together they form a unique fingerprint.

Cite this