PMID: 2110478Feb 13, 1990Paper

A comparison of strategies to stabilize immunoglobulin Fv-fragments

Biochemistry
R GlockshuberA Plückthun

Abstract

Fv-Fragments of antibodies may dissociate at low protein concentrations and are too unstable for many applications at physiological temperatures. To stabilize Fv-fragments against dissociation, we have tested and compared three different strategies on the Fv-fragment of the well-characterized phosphocholine binding antibody McPC603 expressed and secreted in Escherichia coli: chemical cross-linking of the variable domains, introduction of an intermolecular disulfide bond, and construction of a peptide linker to produce a "single-chain" Fv-fragment. All the linked fragments show hapten affinities nearly identical with that of the whole antibody independent of protein concentration and are significantly (up to 60-fold) stabilized against irreversible thermal denaturation. All genetically engineered linked Fv-fragments can be obtained in native conformation in E. coli. The reported strategies for generating Fv-fragments with improved physicochemical properties may extend their usefulness in biotechnology as well as in therapeutic and diagnostic applications.

References

Jun 15, 1976·Biochemistry·J HochmanD Givol
Aug 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·J S HustonR Crea
Oct 28, 1988·Science·J Kraut
Oct 1, 1988·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·R A Lerner, S J Benkovic
Aug 20, 1986·Journal of Molecular Biology·Y SatowD R Davies
Oct 21, 1988·Science·R E BirdM Whitlow
Jan 1, 1987·Methods in Enzymology·J Vieira, J Messing
Jan 1, 1987·Methods in Enzymology·T A KunkelR A Zakour
Jan 1, 1987·Cold Spring Harbor Symposia on Quantitative Biology·A PlückthunJ Stadlmüller
Jan 1, 1986·Methods in Enzymology·R Jaenicke, R Rudolph
Oct 7, 1986·Biochemistry·C O Pabo, E G Suchanek
Nov 1, 1974·Proceedings of the National Academy of Sciences of the United States of America·D M SegalD R Davies
Dec 1, 1972·Proceedings of the National Academy of Sciences of the United States of America·S RudikoffD R Davies
Feb 29, 1972·Biochemistry·B Chesebro, H Metzger
Apr 30, 1982·Science·A M Lesk, K D Hardman

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Citations

Aug 10, 2000·Journal of Molecular Recognition : JMR·A Skerra
Jan 1, 1997·Protein Science : a Publication of the Protein Society·E C OhageB Steipe
Apr 1, 1995·Molecular Biotechnology·R Rapley
Feb 10, 1994·Journal of Immunological Methods·R J Owens, R J Young
Dec 28, 1994·Journal of Immunological Methods·B AkerströmL Björck
May 11, 1995·Journal of Immunological Methods·U BrinkmannI Pastan
Apr 1, 1991·Molecular Immunology·M Ornatowska, J A Glasel
Oct 1, 1992·Molecular Immunology·J AnthonyS C Ng
Apr 1, 1991·Trends in Biotechnology·R E Bird, B W Walker
May 27, 1994·Biochimica Et Biophysica Acta·U Brinkmann, I Pastan
Dec 1, 1995·Toxicology Letters·P R Pentel, D E Keyler
Jan 12, 1996·Journal of Chromatography. B, Biomedical Applications·L M Martin
Jan 1, 1994·Biotechnology Advances·M LittleS Dübel
Apr 1, 1991·Current Opinion in Immunology·A C Hiatt
Apr 1, 1993·Current Opinion in Immunology·A Skerra
Apr 1, 1991·Current Opinion in Biotechnology·A Plückthun
Aug 1, 1995·Current Opinion in Structural Biology·B Nilsson
Oct 1, 1996·Molecular Medicine Today·U Brinkmann
Jun 1, 2005·Protein Expression and Purification·Fatemeh RahbarizadehAbdol-Amir Allameh
Dec 24, 2003·Trends in Biotechnology·Michael Hust, Stefan Dübel
Nov 1, 2002·Biochemical and Biophysical Research Communications·Kim Bak JensenPeter Kristensen
Aug 26, 1998·FEBS Letters·K M MüllerA Plückthun
Sep 13, 2003·Journal of Immunological Methods·Hiroshi UedaTeruyuki Nagamune
Jan 15, 1997·Journal of Immunological Methods·S M KipriyanovM Little
Oct 6, 1998·Journal of Immunological Methods·R VermaA J George

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