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Thromb Haemost 2009; 101(06): 1012-1019
DOI: 10.1160/TH08-12-0816
Theme Issue Article
Schattauer GmbH

Single-chain antibodies as diagnostic tools and therapeutic agents

Christoph E. Hagemeyer
1   Atherothrombosis and Vascular Biology, Baker IDI Heart and Diabetes Institute, Melbourne, Victoria, Australia
,
Constantin von zur Muhlen
2   Department of Cardiology and Angiology, University of Freiburg, Freiburg, Germany
,
Dominik von Elverfeldt
3   Department of Radiology/Medical Physics, University of Freiburg, Freiburg, Germany
,
Karlheinz Peter
1   Atherothrombosis and Vascular Biology, Baker IDI Heart and Diabetes Institute, Melbourne, Victoria, Australia
› Institutsangaben
Weitere Informationen

Publikationsverlauf

Received: 15. Dezember 2008

Accepted: 13. Februar 2009

Publikationsdatum:
24. November 2017 (online)

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Summary

Over three decades after the generation of the first mouse monoclonal antibodies by Kohler and Milstein, recombinant antibodies are the fastest growing class of therapeutic proteins. Furthermore, antibodies are key detection reagents in research and diagnostics. Technology improvements have provided several approaches to manufacturing human antibodies with high affinity for biologically relevant targets. Approximately 300 development programs for therapeutic antibodies have been reported in industrial and academic laboratories, and this clearly demonstrates the expectations towards antibody technology. Antibody fragments are a subclass with growing clinical importance. This review focuses on single-chain antibodies as one of the smallest possible format for recombinant antibodies and their use as diagnostic tools and therapeutic agents. We describe the structure, selection and production of single-chain antibodies. Furthermore, we review current applications of antibody fragments focusing on thrombus targeting using fibrin- and platelet-specific single-chain antibodies as well as describing novel noninvasive imaging approaches for the diagnosis of thrombosis and inflammation.

Keywords

Antibody fragments - imaging - thrombosis - inflammation - phage-display
 

  • References

  • 1 Kohler G, Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity.. Nature 1975; 256: 495-497.
    CrossrefPubMedGoogle Scholar
  • 2 Skerra A, Pluckthun A. Assembly of a functional immunoglobulin Fv fragment in Escherichia coli.. Science 1988; 240: 1038-1041.
    CrossrefPubMedGoogle Scholar
  • 3 Toleikis L, Broders O, Dubel S. Cloning single-chain antibody fragments (scFv) from hybridoma cells.. Methods Mol Med 2004; 94: 447-458.
    PubMedGoogle Scholar
  • 4 Kipriyanov SM, Moldenhauer G, Schuhmacher J. et al. Bispecific tandem diabody for tumor therapy with improved antigen binding and pharmacokinetics.. J Mol Biol 1999; 293: 41-56.
    CrossrefPubMedGoogle Scholar
  • 5 Hoogenboom HR. Selecting and screening recombinant antibody libraries.. Nat Biotechnol 2005; 23: 1105-1116.
    CrossrefPubMedGoogle Scholar
  • 6 Lonberg N. Human antibodies from transgenic animals.. Nat Biotechnol 2005; 23: 1117-1125.
    CrossrefPubMedGoogle Scholar
  • 7 Jostock T, Dubel S. Screening of molecular repertoires by microbial surface display.. Comb Chem High Throughput Screen 2005; 8: 127-133.
    CrossrefPubMedGoogle Scholar
  • 8 Eisenhardt SU, Schwarz M, Bassler N. et al. Subtractive single-chain antibody (scFv) phage-display: tailoring phage-display for high specificity against function-specific conformations of cell membrane molecules.. Nat Protoc 2007; 2: 3063-3073.
    CrossrefPubMedGoogle Scholar
  • 9 Robert R, Jacobin-Valat MJ, Daret D. et al. Identification of human scFvs targeting atherosclerotic lesions: selection by single round in vivo phage display.. J Biol Chem 2006; 281: 40135-40143.
    CrossrefPubMedGoogle Scholar
  • 10 Valadon P, Garnett JD, Testa JE. et al. Screening phage display libraries for organ-specific vascular immunotargeting in vivo.. Proc Natl Acad Sci USA 2006; 103: 407-412.
    CrossrefPubMedGoogle Scholar
  • 11 Todorovska A, Roovers RC, Dolezal O. et al. Design and application of diabodies, triabodies and tetra-bodies for cancer targeting.. J Immunol Methods 2001; 248: 47-66.
    CrossrefPubMedGoogle Scholar
  • 12 Simmons LC, Reilly D, Klimowski L. et al. Expression of full-length immunoglobulins in Escherichia coli: rapid and efficient production of aglycosylated antibodies.. J Immunol Methods 2002; 263: 133-147.
    CrossrefPubMedGoogle Scholar
  • 13 Backmann N, Zahnd C, Huber F. et al. A label-free immunosensor array using single-chain antibody fragments.. Proc Natl Acad Sci USA 2005; 102: 14587-14592.
    CrossrefPubMedGoogle Scholar
  • 14 Shen Z, Mernaugh RL, Yan H. et al. Engineered recombinant single-chain fragment variable antibody for immunosensors.. Anal Chem 2005; 77: 6834-6842.
    CrossrefPubMedGoogle Scholar
  • 15 Shen Z, Stryker GA, Mernaugh RL. et al. Single-chain fragment variable antibody piezoimmunosensors.. Anal Chem 2005; 77: 797-805.
    CrossrefPubMedGoogle Scholar
  • 16 Shen Z, Yan H, Zhang Y. et al. Engineering peptide linkers for scFv immunosensors.. Anal Chem 2008; 80: 1910-1917.
    CrossrefPubMedGoogle Scholar
  • 17 Dietrich T, Perlitz C, Licha K. et al. ED-B fibronectin (ED-B) can be targeted using a novel single chain antibody conjugate and is associated with macrophage accumulation in atherosclerotic lesions.. Basic Res Cardiol 2007; 102: 298-307.
    CrossrefPubMedGoogle Scholar
  • 18 Gawaz M, Langer H, May AE. Platelets in inflammation and atherogenesis.. J Clin Invest 2005; 115: 3378-3384.
    CrossrefPubMedGoogle Scholar
  • 19 Gawaz M, Stellos K, Langer HF. Platelets modulate atherogenesis and progression of atherosclerotic plaques via interaction with progenitor and dendritic cells.. J Thromb Haemost 2008; 6: 235-242.
    CrossrefPubMedGoogle Scholar
  • 20 Schwarz M, Katagiri Y, Kotani M. et al. Reversibility versus persistence of GPIIb/IIIa blocker-induced conformational change of GPIIb/IIIa (alphaIIbbeta3, CD41/CD61).. J Pharmacol Exp Ther 2004; 308: 1002-1011.
    PubMedGoogle Scholar
  • 21 McAteer MA, Schneider JE, Ali ZA. et al. Magnetic resonance imaging of endothelial adhesion molecules in mouse atherosclerosis using dual-targeted microparticles of iron oxide.. Arterioscler Thromb Vasc Biol 2008; 28: 77-83.
    CrossrefPubMedGoogle Scholar
  • 22 McAteer MA, Sibson NR, von Zur Muhlen C. et al. In vivo magnetic resonance imaging of acute brain inflammation using microparticles of iron oxide.. Nat Med 2007; 13: 1253-1258.
    CrossrefPubMedGoogle Scholar
  • 23 Shapiro EM, Skrtic S, Sharer K. et al. MRI detection of single particles for cellular imaging.. Proc Natl Acad Sci USA 2004; 101: 10901-10906.
    CrossrefPubMedGoogle Scholar
  • 24 von zur Muhlen C, von Elverfeldt D, Moeller JA. et al. Magnetic resonance imaging contrast agent targeted toward activated platelets allows in vivo detection of thrombosis and monitoring of thrombolysis.. Circulation 2008; 118: 258-267.
    CrossrefPubMedGoogle Scholar
  • 25 von Zur Muhlen C, Peter K, Ali ZA. et al. Visualization of Activated Platelets by Targeted Magnetic Resonance Imaging Utilizing Conformation-Specific Antibodies against Glycoprotein IIb/IIIa.. J Vasc Res 2009; 46: 6-14.
    CrossrefPubMedGoogle Scholar
  • 26 von Zur Muhlen C, von Elverfeldt D, Choudhury RP. et al. Functionalized magnetic resonance contrast agent selectively binds to glycoprotein IIb/IIIa on activated human platelets under flow conditions and is detectable at clinically relevant field strengths.. Mol Imaging 2008; 7: 59-67.
    PubMedGoogle Scholar
  • 27 von Zur Muhlen C, Sibson NR, Peter K. et al. A contrast agent recognizing activated platelets reveals murine cerebral malaria pathology undetectable by conventional MRI.. J Clin Invest 2008; 118: 1198-1207.
    PubMedGoogle Scholar
  • 28 Evans MJ, Rollins SA, Wolff DW. et al. In vitro and in vivo inhibition of complement activity by a single-chain Fv fragment recognizing human C5.. Mol Immunol 1995; 32: 1183-1195.
    CrossrefPubMedGoogle Scholar
  • 29 Thomas TC, Rollins SA, Rother RP. et al. Inhibition of complement activity by humanized anti-C5 antibody and single-chain Fv.. Mol Immunol 1996; 33: 1389-1401.
    CrossrefPubMedGoogle Scholar
  • 30 Fitch JC, Rollins S, Matis L. et al. Pharmacology and biological efficacy of a recombinant, humanized, single-chain antibody C5 complement inhibitor in patients undergoing coronary artery bypass graft surgery with cardiopulmonary bypass.. Circulation 1999; 100: 2499-2506.
    CrossrefPubMedGoogle Scholar
  • 31 Theroux P, Armstrong PW, Mahaffey KW. et al. Prognostic significance of blood markers of inflammation in patients with ST-segment elevation myocardial infarction undergoing primary angioplasty and effects of pexelizumab, a C5 inhibitor: a substudy of the COMMA trial.. Eur Heart J 2005; 26: 1964-1970.
    CrossrefPubMedGoogle Scholar
  • 32 Mahaffey KW, Van de Werf F, Shernan SK. et al. Effect of pexelizumab on mortality in patients with acute myocardial infarction or undergoing coronary artery bypass surgery: a systematic overview.. Am Heart J 2006; 152: 291-296.
    CrossrefPubMedGoogle Scholar
  • 33 Casalvilla R, Duenas M, Ayala M. et al. A bacterial single-chain Fv antibody fragment that inhibits binding of its parental anti-E-selectin monoclonal antibody to activated human endothelial cells.. J Biotechnol 1999; 72: 1-12.
    CrossrefPubMedGoogle Scholar
  • 34 Swers JS, Widom A, Phan U. et al. A high affinity human antibody antagonist of P-selectin mediated rolling.. Biochem Biophys Res Commun 2006; 350: 508-513.
    CrossrefPubMedGoogle Scholar
  • 35 Friedman G, Jankowski S, Shahla M. et al. Administration of an antibody to E-selectin in patients with septic shock.. Crit Care Med 1996; 24: 229-233.
    CrossrefPubMedGoogle Scholar
  • 36 Borges E, Eytner R, Moll T. et al. The P-selectin glycoprotein ligand-1 is important for recruitment of neutrophils into inflamed mouse peritoneum.. Blood 1997; 90: 1934-1942.
    PubMedGoogle Scholar
  • 37 Eisenhardt SU, Schwarz M, Schallner N. et al. Generation of activation-specific human anti-alphaMbeta2 single-chain antibodies as potential diagnostic tools and therapeutic agents.. Blood 2007; 109: 3521-3528.
    CrossrefPubMedGoogle Scholar
  • 38 Hui KY, Haber E, Matsueda GR. Monoclonal antibodies to a synthetic fibrin-like peptide bind to human fibrin but not fibrinogen.. Science 1983; 222: 1129-1132.
    CrossrefPubMedGoogle Scholar
  • 39 Holvoet P, Laroche Y, Stassen JM. et al. Pharmacokinetic and thrombolytic properties of chimeric plasminogen activators consisting of a single-chain Fv fragment of a fibrin-specific antibody fused to single-chain urokinase.. Blood 1993; 81: 696-703.
    PubMedGoogle Scholar
  • 40 Yan JP, Ko JH, Qi YP. Generation and characterization of a novel single-chain antibody fragment specific against human fibrin clots from phage display antibody library.. Thromb Res 2004; 114: 205-211.
    CrossrefPubMedGoogle Scholar
  • 41 Peter K, Graeber J, Kipriyanov S. et al. Construction and functional evaluation of a single-chain antibody fusion protein with fibrin targeting and thrombin inhibition after activation by factor Xa.. Circulation 2000; 101: 1158-1164.
    CrossrefPubMedGoogle Scholar
  • 42 Weitz JI, Leslie B, Hudoba M. Thrombin binds to soluble fibrin degradation products where it is protected from inhibition by heparin-antithrombin but susceptible to inactivation by antithrombin-independent inhibitors.. Circulation 1998; 97: 544-552.
    CrossrefPubMedGoogle Scholar
  • 43 Meyer BJ, Badimon JJ, Chesebro JH. et al. Dissolution of mural thrombus by specific thrombin inhibition with r-hirudin: comparison with heparin and aspirin.. Circulation 1998; 97: 681-685.
    CrossrefPubMedGoogle Scholar
  • 44 Hagemeyer CE, Tomic I, Jaminet P. et al. Fibrin-targeted direct factor Xa inhibition: construction and characterization of a recombinant factor Xa inhibitor composed of an anti-fibrin single-chain antibody and tick anticoagulant peptide.. Thromb Haemost 2004; 92: 47-53.
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 45 Waxman L, Smith DE, Arcuri KE. et al. Tick anticoagulant peptide (TAP) is a novel inhibitor of blood coagulation factor Xa.. Science 1990; 248: 593-596.
    CrossrefPubMedGoogle Scholar
  • 46 Sitko GR, Ramjit DR. Stabilito, II, et al. Conjunctive enhancement of enzymatic thrombolysis and prevention of thrombotic reocclusion with the selective factor Xa inhibitor, tick anticoagulant peptide. Comparison to hirudin and heparin in a canine model of acute coronary artery thrombosis.. Circulation 1992; 85: 805-815.
    CrossrefPubMedGoogle Scholar
  • 47 Armstrong PW, Collen D, Antman E. Fibrinolysis for acute myocardial infarction: the future is here and now.. Circulation 2003; 107: 2533-2537.
    CrossrefPubMedGoogle Scholar
  • 48 Dewerchin M, Vandamme AM, Holvoet P. et al. Thrombolytic and pharmacokinetic properties of a recombinant chimeric plasminogen activator consisting of a fibrin fragment D-dimer specific humanized monoclonal antibody and a truncated single-chain urokinase.. Thromb Haemost 1992; 68: 170-179.
    Artikel in Thieme ConnectPubMedGoogle Scholar
  • 49 Hagemeyer CE, Tomic I, Weirich U. et al. Construction and characterization of a recombinant plasminogen activator composed of an anti-fibrin single-chain antibody and low-molecular-weight urokinase.. J Thromb Haemost 2004; 2: 797-803.
    CrossrefPubMedGoogle Scholar
  • 50 Bi Q, Cen X, Huang Y. et al. Construction and characterization of trifunctional single-chain urokinase-type plasminogen activators.. Eur J Biochem 2002; 269: 1708-1713.
    CrossrefPubMedGoogle Scholar
  • 51 Ding FX, Yan HL, Lu YM. et al. Cloning, purification and biochemical characterization of a thrombusditargeting thrombolytic agent, comprised of annexin B1, ScuPA-32K and fibrin-adherent peptide.. J Biotechnol 2006; 126: 394-405.
    CrossrefPubMedGoogle Scholar
  • 52 Danielyan K, Ding BS, Gottstein C. et al. Delivery of anti-platelet-endothelial cell adhesion molecule single-chain variable fragment-urokinase fusion protein to the cerebral vasculature lyses arterial clots and attenuates postischemic brain edema.. J Pharmacol Exp Ther 2007; 321: 947-952.
    CrossrefPubMedGoogle Scholar
  • 53 Ding BS, Dziubla T, Shuvaev VV. et al. Advanced drug delivery systems that target the vascular endothelium.. Mol Interv 2006; 6: 98-112.
    CrossrefPubMedGoogle Scholar
  • 54 Ding BS, Gottstein C, Grunow A. et al. Endothelial targeting of a recombinant construct fusing a PECAM-1 single-chain variable antibody fragment (scFv) with prourokinase facilitates prophylactic thrombolysis in the pulmonary vasculature.. Blood 2005; 106: 4191-4198.
    CrossrefPubMedGoogle Scholar
  • 55 Ding BS, Hong N, Murciano JC. et al. Prophylactic thrombolysis by thrombin-activated latent prourokinase targeted to PECAM-1 in the pulmonary vasculature.. Blood 2008; 111: 1999-2006.
    CrossrefPubMedGoogle Scholar
  • 56 Develter J, Booth NA, Declerck PJ. et al. Bispecific targeting of thrombin activatable fibrinolysis inhibitor and plasminogen activator inhibitor-1 by a heterodimer diabody.. J Thromb Haemost 2008; 6: 1884-1891.
    CrossrefPubMedGoogle Scholar
  • 57 Bhatt DL, Topol EJ. Scientific and therapeutic advances in antiplatelet therapy.. Nat Rev Drug Discov 2003; 2: 15-28.
    CrossrefPubMedGoogle Scholar
  • 58 Schwarz M, Meade G, Stoll P. et al. Conformation-specific blockade of the integrin GPIIb/IIIa: a novel antiplatelet strategy that selectively targets activated platelets.. Circ Res 2006; 99: 25-33.
    CrossrefPubMedGoogle Scholar
  • 59 Schwarz M, Rottgen P, Takada Y. et al. Single-chain antibodies for the conformation-specific blockade of activated platelet integrin alphaIIbbeta3 designed by subtractive selection from naive human phage libraries.. Faseb J 2004; 18: 1704-1706.
    CrossrefPubMedGoogle Scholar
  • 60 Jacobin MJ, Robert R, Pouns O. et al. Improving selection of alphaIIbbeta3-binding phage antibodies with increased reactivity derived from immunized donors.. Clin Immunol 2003; 108: 199-210.
    CrossrefPubMedGoogle Scholar
  • 61 Robert R, Clofent-Sanchez G, Hocquellet A. et al. Large-scale production, bacterial localization assessment and immobilized metal affinity chromatography purification of a human single-chain Fv antibody against alphaIIb-beta3 integrin.. Int J Biol Macromol 2006; 39: 51-59.
    CrossrefPubMedGoogle Scholar
  • 62 Robert R, Noubhani AM, Jacobin MJ. et al. Improvement in production and purification bioprocesses of bacterially expressed anti-alphaIIbbeta3 human single-chain FV antibodies.. J Chromatogr B Analyt Technol Biomed Life Sci 2005; 818: 43-51.
    CrossrefPubMedGoogle Scholar
  • 63 Billheimer JT, Dicker IB, Wynn R. et al. Evidence that thrombocytopenia observed in humans treated with orally bioavailable glycoprotein IIb/IIIa antagonists is immune mediated.. Blood 2002; 99: 3540-3546.
    CrossrefPubMedGoogle Scholar
  • 64 Stoll P, Bassler N, Hagemeyer CE. et al. Targeting ligand-induced binding sites on GPIIb/IIIa via single-chain antibody allows effective anticoagulation without bleeding time prolongation.. Arterioscler Thromb Vasc Biol 2007; 27: 1206-1212.
    CrossrefPubMedGoogle Scholar
  • 65 Hagay Y, Lahav J, Levanon A. et al. Molecular characterization of a human monoclonal antibody that interacts with a sulfated tyrosine-containing epitope of the GPIb receptor and inhibits platelet functions.. Mol Immunol 2006; 43: 443-453.
    CrossrefPubMedGoogle Scholar
  • 66 Zhu H, Wang Y, Jiang M. et al. Generation and characterization of a recombinant single chain Fv antibody to von Willebrand factor A1 domain from phage display library.. Thromb Res 2005; 116: 385-391.
    CrossrefPubMedGoogle Scholar
  • 67 Cardinale A, Filesi I, Mattei S. et al. Intracellular targeting and functional analysis of single-chain Fv fragments in mammalian cells.. Methods 2004; 34: 171-178.
    CrossrefPubMedGoogle Scholar
  • 68 Lobato MN, Rabbitts TH. Intracellular antibodies as specific reagents for functional ablation: future therapeutic molecules.. Curr Mol Med 2004; 4: 519-528.
    CrossrefPubMedGoogle Scholar
  • 69 Strebe N, Guse A, Schungel M. et al. Functional knockdown of VCAM-1 at the posttranslational level with ER retained antibodies.. J Immunol Methods 2009; 341: 30-40.
    CrossrefPubMedGoogle Scholar