Skip to main content Accessibility help

We use cookies to distinguish you from other users and to provide you with a better experience on our websites. Close this message to accept cookies or find out how to manage your cookie settings.

Close cookie message
×
Hostname: page-component-cd9895bd7-gbm5v Total loading time: 0 Render date: 2024-12-21T18:13:46.806Z Has data issue: false hasContentIssue false

16 - Single-Domain Antibodies

from PART VI - NOVEL ANTIBODY FORMATS

Published online by Cambridge University Press:  15 December 2009

By
Serge Muyldermans ,
Gholamreza Hassanzadeh Ghassabeh  and
Dirk Saerens
Edited by
Melvyn Little
Melvyn Little
Affiliation:
Affimed Therapeutics AG
Get access

Summary

The antigen-binding entity of an antibody, reduced in size to one single domain, is referred to as a “single-domain antibody.” Various strategies have been explored with variable success to arrive at functional single-domain antibodies. The potential of single-domain antibodies, as research tools or in medicine, is reflected by the three companies – founded in Europe – with a mission to bring these molecules to the market. Domantis using human VH-derived single-domain antibodies started in 2000 and was bought by GSK for £300M in December 2007. Haptogen employing shark single-domain antibodies was acquired by Wyeth, and Ablynx focusing on llama-derived single-domain antibodies received over €70M in three rounds of venture capitalist investments and another €80M on the Euronext stock market in November 2007. Regarding therapeutic applications, Arana Therapeutics in Australia entered a Phase 2 clinical trial with its single-domain antibody derivative. In this chapter, we will review (1) the various antibodies used for generating single-domain antibodies, (2) the properties of single-domain antibodies that create an added value for use in immunotherapy, and (3) a number of therapeutic applications.

THE DEVELOPMENT OF SINGLE-DOMAIN ANTIBODIES

Antibodies comprise two identical heavy chain polypeptides (H) carrying chains of carbohydrates and two identical light chain proteins (L). Their ability to bind specifically to an antigen is dictated by the paired variable regions of the heavy (VH) and the light (VL) chain (Figure 16.1).

Type
Chapter
Information
Recombinant Antibodies for Immunotherapy , pp. 216 - 230
Publisher: Cambridge University Press
Print publication year: 2009

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Baral, T.N., Magez, S., et al. (2006). Experimental therapy of African trypanosomiasis with a nanobody-conjugated human trypanolytic factor. Nat Med 12, 580–584.CrossRefGoogle ScholarPubMed
Barthelemy, P.A., Raab, H., et al. (2008). Comprehensive analysis of the factors contributing to the stability and solubility of autonomous human VH domains. J Biol Chem 283, 3639–3654.CrossRefGoogle ScholarPubMed
Behar, G., Siberil, S., et al. (2008). Isolation and characterization of anti-FcgammaRIII (CD16) llama single-domain antibodies that activate natural killer cells. Protein Eng Des Sel 21, 1–10.CrossRefGoogle ScholarPubMed
Conrath, K., Vincke, C., et al. (2005). Antigen binding and solubility effects upon the veneering of a camel VHH in framework-2 to mimic a VH. J Mol Biol 350, 112–125.CrossRefGoogle Scholar
Coppieters, K., Dreier, T., et al. (2006). Formatted anti-tumor necrosis factor alpha VHH proteins derived from camelids show superior potency and targeting to inflamed joints in a murine model of collagen-induced arthritis. Arthritis Rheum 54, 1856–1866.CrossRefGoogle Scholar
Cortez-Retamozo, V., Backmann, N., et al. (2004). Efficient cancer therapy with a nanobody-based conjugate. Cancer Res 64, 2853–2857.CrossRefGoogle ScholarPubMed
Cortez-Retamozo, V., Lauwereys, M., et al. (2002). Efficient tumor targeting by single-domain antibody fragments of camels. Int J Cancer 98, 456–462.CrossRefGoogle ScholarPubMed
Davies, J., Riechmann, L. (1995). Antibody VH domains as small recognition units. Biotechnol 13, 475–479.Google ScholarPubMed
Genst, E., Saerens, D., et al. (2006a). Antibody repertoire development in camelids. Dev Comp Immunol 30, 187–198.CrossRefGoogle ScholarPubMed
Genst, E., Silence, K., et al. (2006b). Molecular basis for the preferential cleft recognition by dromedary heavy-chain antibodies. Proc Natl Acad Sci USA 103, 4586–4591.CrossRefGoogle ScholarPubMed
Haard, H.J., Bezemer, S., et al. (2005). Llama antibodies against a lactococcal protein located at the tip of the phage tail prevent phage infection. J Bacteriol 187, 4531–4541.CrossRefGoogle ScholarPubMed
Dekker, S., Toussaint, W., et al. (2003). Intracellularly expressed single-domain antibody against p15 matrix protein prevents the production of porcine retroviruses. J Virol 77, 12132–12139.CrossRefGoogle ScholarPubMed
Diaz, M., Stanfield, R.L., et al. (2002). Structural analysis, selection, and ontogeny of the shark new antigen receptor (IgNAR): identification of a new locus preferentially expressed in early development. Immunogenetics 54, 501–512.CrossRefGoogle ScholarPubMed
Dooley, H., Flajnik, M.F., et al. (2003). Selection and characterization of naturally occurring single-domain (IgNAR) antibody fragments from immunized sharks by phage display. Mol Immunol 40, 25–33.CrossRefGoogle ScholarPubMed
Dumoulin, M., Last, A.M., et al. (2003). A camelid antibody fragment inhibits the formation of amyloid fibrils by human lysozyme. Nature 424, 783–788.CrossRefGoogle ScholarPubMed
Dumoulin, M., Conrath, K., et al. (2002). Single-domain antibody fragments with high conformational stability. Protein Sci 11, 500–515.CrossRefGoogle ScholarPubMed
El Khattabi, M., Adams, H., et al., (2006). Llama single-chain antibody that blocks lipopolysaccharide binding and signaling: prospects for therapeutic applications. Clin Vaccine Immunol 13, 1079–1086.CrossRefGoogle ScholarPubMed
Ferrari, A., Rodriguez, M.M., et al. (2007). Immunobiological role of llama heavy-chain antibodies against a bacterial beta-lactamase. Vet Immunol Immunopathol 117, 173–182.CrossRefGoogle ScholarPubMed
Frenken, L.G., Linden, R.H., et al. (2000). Isolation of antigen specific llama VHH antibody fragments and their high level secretion by Saccharomyces cerevisiae. J Biotechnol 78, 11–21.CrossRefGoogle ScholarPubMed
Goldman, E.R., Anderson, G.P., et al. (2006). Facile generation of heat-stable antiviral and antitoxin single domain antibodies from a semisynthetic llama library. Anal Chem 78, 8245–8255.CrossRefGoogle ScholarPubMed
Haber, E., Richards, F.F. (1966). The specificity of antigenic recognition of antibody heavy chain. Proc R Soc Lond B Biol Sci 166, 176–187.CrossRefGoogle ScholarPubMed
Habicht, G., Haupt, C., et al. (2007) Directed selection of a conformational antibody domain that prevents mature amyloid fibril formation by stabilizing Aβ protofibrils. Proc Natl Acad Sci USA 104, 19232–19237.CrossRefGoogle ScholarPubMed
Harmsen, M.M., Solt, C.B., et al. (2005). Prolonged in vivo residence times of llama single-domain antibody fragments in pigs by binding to porcine immunoglobulins. Vaccine 23, 4926–4934.CrossRefGoogle ScholarPubMed
Harmsen, M.M., Solt, C.B., et al., (2006). Selection and optimization of proteolytically stable llama single-domain antibody fragments for oral immunotherapy. Appl Microbiol Biotechnol 72, 544–551.CrossRefGoogle ScholarPubMed
Harmsen, M.M., Solt, C.B., et al. (2007). Passive immunization of guinea pigs with llama single-domain antibody fragments against foot-and-mouth disease. Vet Microbiol 120, 193–206.CrossRefGoogle ScholarPubMed
Hmila, I., Abdallah, R.B., et al. (2008) VHH, bivalent domains and chimeric heavy chain-only antibodies with high neutralizing efficacy for scorpion toxin AahI. Mol Immunol 45, in press.CrossRefGoogle Scholar
Holt, L.J., Basran, A., et al. (2008). Anti-serum albumin domain antibodies for extending the half-lives of short lived drugs. Protein Eng Des Sel 21, 283–288.CrossRefGoogle ScholarPubMed
Hulstein, J.J., Groot, P.G., et al. (2005). A novel nanobody that detects the gain-of-function phenotype of von Willebrand factor in ADAMTS13 deficiency and von Willebrand disease type 2B. Blood 106, 3035–3042.CrossRefGoogle ScholarPubMed
Ismaili, A., Jalali-Javaran, M., et al. (2007). Production and characterization of anti-(mucin MUC1) single-domain antibody in tobacco (Nicotiana tabacum cultivar Xanthi). Biotechnol Appl Biochem 47, 11–19.Google Scholar
Janssens, R., Dekker, S., et al. (2006). Generation of heavy-chain-only antibodies in mice. Proc Natl Acad Sci USA 103, 15130–15135.CrossRefGoogle ScholarPubMed
Jobling, S.A., Jarman, C., et al. (2003). Immunomodulation of enzyme function in plants by single-domain antibody fragments. Nat Biotechnol 21, 77–80.CrossRefGoogle ScholarPubMed
Koide, A., Bailey, C.W., et al. (1998). The fibronectin type III domain as a scaffold for novel binding proteins. J Mol Biol 284, 1141–1151.CrossRefGoogle ScholarPubMed
Kruger, C., Hultberg, A., et al. (2006). Therapeutic effect of llama derived VHH fragments against Streptococcus mutans on the development of dental caries. Appl Microbiol Biotechnol 72, 732–737.CrossRefGoogle ScholarPubMed
Liu, J.L., Anderson, G.P., et al. (2007a). Selection of cholera toxin specific IgNAR single-domain antibodies from a naive shark library. Mol Immunol 44, 1775–1783.CrossRefGoogle ScholarPubMed
Liu, J.L., Anderson, G.P., et al. (2007b). Isolation of anti-toxin single domain antibodies from a semi-synthetic spiny dogfish shark display library. BMC Biotechnol 7, 78.CrossRefGoogle ScholarPubMed
Miao, Q.F., Liu, X.Y., et al. (2007). An enediyne-energized single-domain antibody-containing fusion protein shows potent antitumor activity. Anticancer Drugs 18, 127–137.CrossRefGoogle ScholarPubMed
Muruganandam, A., Tanha, J., et al. (2002). Selection of phage-displayed llama single-domain antibodies that transmigrate across human blood-brain barrier endothelium. Faseb J 16, 240–242.CrossRefGoogle ScholarPubMed
Nuttall, S.D., Krishnan, U.V., et al. (2003). Isolation and characterization of an IgNAR variable domain specific for the human mitochondrial translocase receptor Tom70. Eur J Biochem 270, 3543–3554.CrossRefGoogle ScholarPubMed
Nguyen, V.K., Su, C., et al. (2002). Heavy-chain antibodies in Camelidae; a case of evolutionary innovation. Immunogenetics 54, 39–47.Google ScholarPubMed
Olichon, A., Schweizer, D., et al. (2007). Heating as a rapid purification method for recovering correctly-folded thermotolerant VH and VHH domains. BMC Biotechnol 7, 7.CrossRefGoogle ScholarPubMed
Pant, N., Hultberg, A., et al. (2006). Lactobacilli expressing variable domain of llama heavy-chain antibody fragments (lactobodies) confer protection against rotavirus-induced diarrhea. J Infect Dis 194, 1580–1588.CrossRefGoogle ScholarPubMed
Paz, K., Brennan, L.A., et al. (2005). Human single-domain neutralizing intrabodies directed against Etk kinase: a novel approach to impair cellular transformation. Mol Cancer Ther 4, 1801–1809.CrossRefGoogle ScholarPubMed
Pessi, A., Bianchi, E., et al. (1993). A designed metal-binding protein with a novel fold. Nature 362, 367–369.CrossRefGoogle ScholarPubMed
Rahbarizadeh, F., Rasaee, M.J., et al. (2006). Over expression of anti-MUC1 single-domain antibody fragments in the yeast Pichia pastoris. Mol Immunol 43, 426–435.CrossRefGoogle ScholarPubMed
Reiter, Y., Schuck, P., et al. (1999). An antibody single-domain phage display library of a native heavy chain variable region: isolation of functional single-domain VH molecules with a unique interface. J Mol Biol 290, 685–698.CrossRefGoogle ScholarPubMed
Roovers, R.C., Laeremans, T., et al. (2007). Efficient inhibition of EGFR signaling and of tumour growth by antagonistic anti-EFGR nanobodies. Cancer Immunol Immunother 56, 303–317.CrossRefGoogle ScholarPubMed
Rothbauer, U., Zolghadr, K., et al. (2008). A versatile nanotrap for biochemical and functional studies with fluorescent fusion proteins. Mol Cell Proteomics 7, 282–289.CrossRefGoogle ScholarPubMed
Saerens, D., Stijlemans, B., et al. (2008a). Parallel selection of multiple anti-infectome nanobodies without access to purified antigens. J Immunol Methods 329, 138–150.CrossRefGoogle ScholarPubMed
Saerens, D., Conrath, K., et al. (2008b). Disulfide bond introduction for general stabilization of immunoglobulin heavy-chain variable domains. J Mol Biol 377, 478–488.CrossRefGoogle ScholarPubMed
Shao, C.Y., Secombes, C.J., et al. (2007). Rapid isolation of IgNAR variable single-domain antibody fragments from a shark synthetic library. Mol Immunol 44, 656–665.CrossRefGoogle ScholarPubMed
Stanfield, R.L., Dooley, H., et al. (2004). Crystal structure of a shark single-domain antibody V region in complex with lysozyme. Science 305, 1770–1773.CrossRefGoogle ScholarPubMed
Stewart, C.S., MacKenzie, C.R., et al. (2007). Isolation, characterization and pentamerization of alpha-cobrotoxin specific single-domain antibodies from a naive phage display library: preliminary findings for antivenom development. Toxicon 49, 699–709.CrossRefGoogle ScholarPubMed
Streltsov, V.A., Carmichael, J.A., et al. (2005). Structure of a shark IgNAR antibody variable domain and modeling of an early-developmental isotype. Protein Sci 14, 2901–2909.CrossRefGoogle ScholarPubMed
Szynol, A., Haard, J.J., et al. (2006). Design of a peptibody consisting of the antimicrobial peptide dhvar5 and a llama variable heavy-chain antibody fragment. Chem Biol Drug Des 67, 425–431.CrossRefGoogle Scholar
Thomassen, Y.E., Meijer, W., et al. (2002). Large-scale production of VHH antibody fragments by Saccharomyces cerevisiae. Enzyme Microb. Technol. 30, 273–278.CrossRefGoogle Scholar
Utsumi, S., Karush, F. (1964). The Subunits of Purified Rabbit Antibody. Biochemistry 3, 1329–1338.CrossRefGoogle ScholarPubMed
Linden, R.H., Geus, B., et al. (2000). Improved production and function of llama heavy chain antibody fragments by molecular evolution. J Biotechnol 80, 261–270.CrossRefGoogle ScholarPubMed
Linden, R.H., Frenken, L.G., et al. (1999). Comparison of physical chemical properties of llama VHH antibody fragments and mouse monoclonal antibodies. Biochim Biophys Acta 1431, 37–46.CrossRefGoogle ScholarPubMed
Vaart, JM., Pant, N., et al. (2006). Reduction in morbidity of rotavirus induced diarrhoea in mice by yeast produced monovalent llama-derived antibody fragments. Vaccine 24, 4130–4137.Google ScholarPubMed
Koningsbruggen, S., Haard, H., et al. (2003). Llama-derived phage display antibodies in the dissection of the human disease oculopharyngeal muscular dystrophy. J Immunol Methods 279, 149–161.CrossRefGoogle ScholarPubMed
Verheesen, P., Kluijver, A., et al. (2006a). Prevention of oculopharyngeal muscular dystrophy-associated aggregation of nuclear polyA-binding protein with a single-domain intracellular antibody. Hum Mol Genet 15, 105–111.CrossRefGoogle ScholarPubMed
Verheesen, P., Roussis, A., et al. (2006b). Reliable and controllable antibody fragment selections from Camelid non-immune libraries for target validation. Biochim Biophys Acta 1764, 1307–1319.CrossRefGoogle ScholarPubMed
Ward, E.S., Gussow, D., et al. (1989). Binding activities of a repertoire of single immunoglobulin variable domains secreted from Escherichia coli. Nature 341, 544–546.CrossRefGoogle ScholarPubMed
Worn, A., Pluckthun, A. (2001). Stability engineering of antibody single-chain Fv fragments. J Mol Biol 305, 989–1010.CrossRefGoogle ScholarPubMed
Yau, K.Y., Groves, M.A., et al. (2003). Selection of hapten-specific single-domain antibodies from a non-immunized llama ribosome display library. J Immunol Methods 281, 161–175.CrossRefGoogle ScholarPubMed
Zhang, J., Tanha, J., et al. (2004). Pentamerization of single-domain antibodies from phage libraries: a novel strategy for the rapid generation of high-avidity antibody reagents. J Mol Biol 335, 49–56.CrossRefGoogle ScholarPubMed
Zou, X., Osborn, M.J., et al. (2007) Heavy chain-only antibodies are spontaneously produced in light chain-deficient mice. J Exp Med 204, 3271–3283.CrossRefGoogle ScholarPubMed

Save book to Kindle

To save this book to your Kindle, first ensure [email protected] is added to your Approved Personal Document E-mail List under your Personal Document Settings on the Manage Your Content and Devices page of your Amazon account. Then enter the ‘name’ part of your Kindle email address below. Find out more about saving to your Kindle.

Note you can select to save to either the @free.kindle.com or @kindle.com variations. ‘@free.kindle.com’ emails are free but can only be saved to your device when it is connected to wi-fi. ‘@kindle.com’ emails can be delivered even when you are not connected to wi-fi, but note that service fees apply.

Find out more about the Kindle Personal Document Service.

Available formats
×

Save book to Dropbox

To save content items to your account, please confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your account. Find out more about saving content to Dropbox.

Available formats
×

Save book to Google Drive

To save content items to your account, please confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your account. Find out more about saving content to Google Drive.

Available formats
×