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Engineering antibody fragments to fold in the absence of disulfide bonds
Article first published online: 2 DEC 2008
DOI: 10.1002/pro.31
Copyright © 2008 The Protein Society
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How to Cite
Seo, M. J., Jeong, K. J., Leysath, C. E., Ellington, A. D., Iverson, B. L. and Georgiou, G. (2009), Engineering antibody fragments to fold in the absence of disulfide bonds. Protein Science, 18: 259–267. doi: 10.1002/pro.31
Publication History
- Issue published online: 30 JAN 2009
- Article first published online: 2 DEC 2008
- Accepted manuscript online: 2 DEC 2008 12:00AM EST
- Manuscript Accepted: 2 NOV 2008
- Manuscript Revised: 29 OCT 2008
- Manuscript Received: 5 SEP 2008
Funded by
- ONR
- DARPA
References
- 1, , , ( 1998) Antibody scFv fragments without disulfide bonds made by molecular evolution. J Mol Biol 275: 245–253.
- 2, ( 1998) Mutual stabilization of VL and VH in single-chain antibody fragments, investigated with mutants engineered for stability. Biochemistry 37: 13120–13127.
- 3, , ( 2007) Stabilization of an immunoglobulin fold domain by an engineered disulfide bond at the buried hydrophobic region. J Biol Chem 282: 36489–36495.
- 4, ( 1999) The selection of intracellular antibodies. Trends Biotechnol 17: 115–121.
- 5, , ( 2008) Intracellular antibodies (intrabodies) and their therapeutic potential. Handb Exp Pharmacol 181: 343–373.
- 6, ( 2006) The therapeutic potential of intrabodies in neurologic disorders: focus on Huntington and Parkinson diseases. Bio Drugs 20: 327–333.
- 7, ( 2006) Intrabody-based approaches to cancer therapy: status and prospects. Curr Med Chem 13: 1473–1480.
- 8, ( 2008) Combating protein misfolding and aggregation by intracellular antibodies. Curr Mol Med 8: 2–11.
- 9, , ( 1997) A natural antibody missing a cysteine in VH: consequences for thermodynamic stability and folding. J Mol Biol 265: 161–172.
- 10, , , , , , , , ( 1999) A single-chain antibody fragment is functionally expressed in the cytoplasm of both Escherichia coli and transgenic plants. Eur J Biochem 262: 617–624.Direct Link:
- 11, , , , ( 1999) Selection of antibodies for intracellular function using a two-hybrid in vivo system. Proc Natl Acad Sci USA 96: 11723–11728.
- 12, , , , ( 2001) Tailoring in vitro evolution for protein affinity or stability. Proc Natl Acad Sci USA 98: 75–80.
- 13, , , , , , , , ( 2002) Direct in vivo screening of intrabody libraries constructed on a highly stable single-chain framework. J Biol Chem 277: 45075–45085.
- 14, , , ( 2005) Design and selection of an intrabody library produced de-novo for the non-structural protein NSP5 of rotavirus. J Immunol Methods 301: 31–40.
- 15, , , ( 2006) Direct selection of antibodies from complex libraries with the protein fragment complementation assay. J Mol Biol 357: 427–441.
- 16, , , ( 2008) In vivo selection of intrabodies specifically targeting protein-protein interactions: a general platform for an “undruggable” class of disease targets. J Biotechnol 135: 1–15.
- 17, , , , , ( 2004) Direct phage to intrabody screening (DPIS): demonstration by isolation of cytosolic intrabodies against the TES1 site of Epstein Barr virus latent membrane protein 1 (LMP1) that block NF-kappaB transactivation. J Mol Biol 335: 193–207.
- 18, , , , , , , , ( 2001) A semi-synthetic repertoire of intrinsically stable antibody fragments derived from a single-framework scaffold. J Mol Biol 310: 603–615.
- 19, , , , , , ( 2003) Engineering stable cytoplasmic intrabodies with designed specificity. J Mol Biol 330: 323–332.
- 20, , , , , , , , , ( 2007) A focused antibody library for selecting scFvs expressed at high levels in the cytoplasm. BMC Biotechnol 7: 81.
- 21, , , , ( 2005) A universal strategy for stable intracellular antibodies. J Immunol Methods 303: 19–39.
- 22, ( 2001) Roles of thiol-redox pathways in bacteria. Annu Rev Microbiol 55: 21–48.
- 23, ( 2004) Catalysis of disulfide bond formation and isomerization in the Escherichia coli periplasm. Biochim Biophys Acta 1694: 111–119.
- 24, ( 2004) Protein thiol modifications visualized in vivo. PLoS Biol 2: e333.
- 25, , , , ( 2008) Laboratory evolution of Escherichia coli thioredoxin for enhanced catalysis of protein oxidation in the periplasm reveals a phylogenetically conserved substrate specificity determinant. J Biol Chem 283: 840–848.
- 26( 2003) Anchored Periplasmic Expression (APEx): a versatile technology for the flow cytometric selection of high affinity antibodies from Eshcerichia coli expressed libraries. Ph.D. Thesis. Austin: The University of Texas, pp 10–14.
- 27, , , , , ( 2004) A periplasmic fluorescent reporter protein and its application in high-throughput membrane protein topology analysis. J Mol Biol 341: 901–909.
- 28, , , ( 2007) APEx 2-hybrid, a quantitative protein-protein interaction assay for antibody discovery and engineering. Proc Natl Acad Sci USA 104: 8247–8252.
- 29, , , , ( 2007) Isolation of engineered, full-length antibodies from libraries expressed in Escherichia coli. Nat Biotechnol 25: 563–565.
- 30, , ( 1988) Production and characterization of monoclonal antibodies to the protective antigen component of Bacillus anthracis toxin. Infect Immun 56: 1807–1813.
- 31, , ( 2003) Expression and purification of the Bacillus anthracis protective antigen domain 4. Protein Expr Purif 27: 325–330.
- 32, ( 1997) In vitro and in vivo redox states of the Escherichia coli periplasmic oxidoreductases DsbA and DsbC. Biochemistry 36: 10067–10072.
- 33, , , ( 2005) Why high-error-rate random mutagenesis libraries are enriched in functional and improved proteins. J Mol Biol 350: 806–816.
- 34, , , ( 1995) Substitution of charged residues into the hydrophobic core of Escherichia coli thioredoxin results in a change in heat capacity of the native protein. Biochemistry 34: 2148–2152.
- 35, ( 2008) Functional intracellular antibody fragments do not require invariant intra-domain disulfide bonds. J Mol Biol 376: 749–757.
- 36, , . ( 2007) A scFv antibody mutant isolated in a genetic screen for improved export via the twin arginine transporter pathway exhibits faster folding. J Mol Biol 369: 631–639.
- 37, , , , , ( 2004) Anchored periplasmic expression, a versatile technology for the isolation of high-affinity antibodies from Escherichia coli-expressed libraries. Proc Natl Acad Sci USA 101: 9193–9198.
- 38, , , ( 2007) Binding and enrichment of Escherichia coli spheroplasts expressing inner membrane tethered scFv antibodies on surface immobilized antigens. Biotechnol Bioeng 98: 39–47.Direct Link:
- 39, , , , , ( 2005) Evolution of highly active enzymes by homology-independent recombination. Proc Natl Acad Sci USA 102: 10082–10087.
- 40, ( 1985) Internal protein motions, concentrated glycerol, and hydrogen exchange studied in myoglobin. Biochemistry 24: 2095–2100.
- 41, , , ( 2004) Engineered DsbC chimeras catalyze both protein oxidation and disulfide-bond isomerization in Escherichia coli: reconciling two competing pathways. Proc Natl Acad Sci USA 101: 10018–10023.
- 42, , , , , ( 2003) Isolation and expression of recombinant antibody fragments to the biological warfare pathogen Brucella melitensis. J Immunol Methods 276: 185–196.

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