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 language | English (US) |
|---|---|
| Pages (from-to) | 67-75 |
| Number of pages | 9 |
| Journal | Journal of Immunological Methods |
| Volume | 201 |
| Issue number | 1 |
| DOIs | |
| State | Published - 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.
Keywords
- CD7
- Pichia pastoris
- desipramine
- single chain Fv
- surface plasmon resonance