Antigen Recognition and Targeted Delivery by the Single-chain Fv

Overview

Journal Cell Biophys

Specialty Biophysics

Date 1993 Jan 1

PMID 7889539

Citations 7

Authors

J S Huston

M S Tai

J McCartney

P Keck

H Oppermann

Affiliations

Soon will be listed here.

Abstract

The single-chain Fv (sFv) has proven attractive for immuno-targeting, both alone and as a targeting element within sFv fusion proteins. This chapter summarizes the features of sFv proteins that have sparked this interest, starting with the conservation of Fv architecture that makes general sFv design practical. The length and composition of linkers used to bridge V domains are discussed based on the sFv literature; special emphasis is given to the (Gly4Ser)3 15-residue linker that has proven of broad utility for constructing Fv regions of antibodies and other members of the immunoglobulin superfamily. The refolding properties of sFv proteins are summarized and examples given from our laboratory. Spontaneous refolding from the fully reduced and denatured state, typified by 26-10 sFv, is contrasted with disulfide-restricted refolding, exemplified by MOPC 315 and R11D10 sFv proteins, which recover antigen binding only if their disulfides have been oxidized prior to removal of denaturant. The medical value of sFv proteins hinges on their reliability in antigen recognition and rapidity in targeted delivery. Detailed analysis of specificity and affinity of antigen binding by the 26-10 antidigoxin sFv has demonstrated very high fidelity to the binding properties of the parent 26-10 sFv. These results gave confidence to the pursuit of more complex biomedical applications of these proteins, which is indicated by our work with the R11D10 sFv for the imaging of myocardial infarctions. Diagnostic imaging and therapeutic immunotargeting by sFv present significant opportunities, particularly as a result of their pharmacokinetic properties. Intravenously administered sFv offers much faster clearance than conventional Fab fragments or intact immunoglobulin with minimal background binding.

Citing Articles

Oral immunization with rotavirus VP7-CTB fusion expressed in transgenic induces antigen-specific IgA and IgG and passive protection in mice.

Li Y, Guan L, Liu X, Liu W, Yang J, Zhang X Exp Ther Med. 2018; 15(6):4866-4874.

PMID: 29805507 PMC: 5952079. DOI: 10.3892/etm.2018.6003.

Single-Chain Soluble BG505.SOSIP gp140 Trimers as Structural and Antigenic Mimics of Mature Closed HIV-1 Env.

Georgiev I, Joyce M, Yang Y, Sastry M, Zhang B, Baxa U J Virol. 2015; 89(10):5318-29.

PMID: 25740988 PMC: 4442528. DOI: 10.1128/JVI.03451-14.

A fibrinogen-derived peptide provides intercellular adhesion molecule-1-specific targeting and intraendothelial transport of polymer nanocarriers in human cell cultures and mice.

Garnacho C, Serrano D, Muro S J Pharmacol Exp Ther. 2011; 340(3):638-47.

PMID: 22160267 PMC: 3286316. DOI: 10.1124/jpet.111.185579.

Ribosome-inactivating proteins: from plant defense to tumor attack.

de Virgilio M, Lombardi A, Caliandro R, Fabbrini M Toxins (Basel). 2011; 2(11):2699-737.

PMID: 22069572 PMC: 3153179. DOI: 10.3390/toxins2112699.

Enhancement of cell type specificity by quantitative modulation of a chimeric ligand.

Cironi P, Swinburne I, Silver P J Biol Chem. 2008; 283(13):8469-76.

PMID: 18230610 PMC: 2417160. DOI: 10.1074/jbc.M708502200.

References

Glockshuber R, Schmidt T, Pluckthun A . The disulfide bonds in antibody variable domains: effects on stability, folding in vitro, and functional expression in Escherichia coli. Biochemistry. 1992; 31(5):1270-9. DOI: 10.1021/bi00120a002. View

Glockshuber R, Malia M, Pfitzinger I, Pluckthun A . A comparison of strategies to stabilize immunoglobulin Fv-fragments. Biochemistry. 1990; 29(6):1362-7. DOI: 10.1021/bi00458a002. View

Ward E . Secretion of T cell receptor fragments from recombinant Escherichia coli cells. J Mol Biol. 1992; 224(4):885-90. DOI: 10.1016/0022-2836(92)90455-s. View

Yokota T, Milenic D, Whitlow M, Schlom J . Rapid tumor penetration of a single-chain Fv and comparison with other immunoglobulin forms. Cancer Res. 1992; 52(12):3402-8. View

Herron J, He X, MASON M, Voss Jr E, EDMUNDSON A . Three-dimensional structure of a fluorescein-Fab complex crystallized in 2-methyl-2,4-pentanediol. Proteins. 1989; 5(4):271-80. DOI: 10.1002/prot.340050404. View

Marquart M, Deisenhofer J, Huber R, PALM W . Crystallographic refinement and atomic models of the intact immunoglobulin molecule Kol and its antigen-binding fragment at 3.0 A and 1.0 A resolution. J Mol Biol. 1980; 141(4):369-91. DOI: 10.1016/0022-2836(80)90252-1. View

Gibbs R, Posner B, Filpula D, Dodd S, Finkelman M, Lee T . Construction and characterization of a single-chain catalytic antibody. Proc Natl Acad Sci U S A. 1991; 88(9):4001-4. PMC: 51581. DOI: 10.1073/pnas.88.9.4001. View

POLJAK R, Amzel L, AVEY H, Chen B, Phizackerley R, Saul F . Three-dimensional structure of the Fab' fragment of a human immunoglobulin at 2,8-A resolution. Proc Natl Acad Sci U S A. 1973; 70(12):3305-10. PMC: 427225. DOI: 10.1073/pnas.70.12.3305. View

Chaudhary V, Queen C, Junghans R, Waldmann T, Fitzgerald D, Pastan I . A recombinant immunotoxin consisting of two antibody variable domains fused to Pseudomonas exotoxin. Nature. 1989; 339(6223):394-7. DOI: 10.1038/339394a0. View

10.

Pack P, Pluckthun A . Miniantibodies: use of amphipathic helices to produce functional, flexibly linked dimeric FV fragments with high avidity in Escherichia coli. Biochemistry. 1992; 31(6):1579-84. DOI: 10.1021/bi00121a001. View

11.

Breitling F, Dubel S, Seehaus T, Klewinghaus I, Little M . A surface expression vector for antibody screening. Gene. 1991; 104(2):147-53. DOI: 10.1016/0378-1119(91)90244-6. View

12.

Marks J, Hoogenboom H, Bonnert T, McCafferty J, Griffiths A, Winter G . By-passing immunization. Human antibodies from V-gene libraries displayed on phage. J Mol Biol. 1991; 222(3):581-97. DOI: 10.1016/0022-2836(91)90498-u. View

13.

Huston J, Levinson D, Tai M, Novotny J, Margolies M, Ridge R . Protein engineering of antibody binding sites: recovery of specific activity in an anti-digoxin single-chain Fv analogue produced in Escherichia coli. Proc Natl Acad Sci U S A. 1988; 85(16):5879-83. PMC: 281868. DOI: 10.1073/pnas.85.16.5879. View

14.

Batra J, Chaudhary V, Fitzgerald D, Pastan I . TGF alpha-anti-Tac(Fv)-PE40: a bifunctional toxin cytotoxic for cells with EGF or IL2 receptors. Biochem Biophys Res Commun. 1990; 171(1):1-6. DOI: 10.1016/0006-291x(90)91347-u. View

15.

Chothia C, Novotny J, Bruccoleri R, Karplus M . Domain association in immunoglobulin molecules. The packing of variable domains. J Mol Biol. 1985; 186(3):651-63. DOI: 10.1016/0022-2836(85)90137-8. View

16.

Sharon J, Givol D . Preparation of Fv fragment from the mouse myeloma XRPC-25 immunoglobulin possessing anti-dinitrophenyl activity. Biochemistry. 1976; 15(7):1591-4. DOI: 10.1021/bi00652a033. View

17.

McCartney J, Lederman L, Drier E, Wu G, Batorsky R, Huston J . Biosynthetic antibody binding sites: development of a single-chain Fv model based on antidinitrophenol IgA myeloma MOPC 315. J Protein Chem. 1991; 10(6):669-83. DOI: 10.1007/BF01025718. View

18.

Khaw B, Fallon F, Strauss H, Haber E . Myocardial infarct imaging of antibodies to canine cardiac myosin with indium-111-diethylenetriamine pentaacetic acid. Science. 1980; 209(4453):295-7. DOI: 10.1126/science.7384803. View

19.

Greer J . Comparative modeling of homologous proteins. Methods Enzymol. 1991; 202:239-52. DOI: 10.1016/0076-6879(91)02014-z. View

20.

Sen J, BEYCHOK S . Proteolytic dissection of a hapten binding site. Proteins. 1986; 1(3):256-62. DOI: 10.1002/prot.340010308. View