Single-domain antibodies as building blocks for novel therapeutics

Current Opinion in Pharmacology

Volumn 8, Issue 5, 2008, Pages 600-608

  Saerens, Dirk ^a,b   Ghassabeh, Gholamreza Hassanzadeh ^a,b   Muyldermans, Serge ^a,b  

^a VRIJE UNIVERSITEIT BRUSSEL   (Belgium)

^b DEPARTMENT OF PLANT SYSTEMS BIOLOGY   (Belgium)

Author keywords

[No Author keywords available]

Indexed keywords

ADALIMUMAB; ALX 0081; ANTIBODY; ANTICOAGULANT AGENT; ART 621; IMMUNOGLOBULIN HEAVY CHAIN; IMMUNOGLOBULIN LIGHT CHAIN; INFLIXIMAB; SCORPION VENOM; TUMOR NECROSIS FACTOR ANTIBODY; UNCLASSIFIED DRUG;

ALZHEIMER DISEASE; ANTIBODY ISOLATION; ANTIGEN BINDING; ANTIGEN SPECIFICITY; CANCER INHIBITION; CLINICAL TRIAL; DENTAL CARIES; DRUG DESIGN; DRUG HALF LIFE; DRUG POTENCY; DRUG SELECTIVITY; DRUG SYNTHESIS; ENVENOMATION; HUMAN; INFECTIOUS DIARRHEA; MOLECULAR SIZE; NONHUMAN; PARKINSON DISEASE; POISONOUS SCORPION; PRIORITY JOURNAL; PROTEIN DOMAIN; PROTEIN ENGINEERING; PSORIASIS VULGARIS; REVIEW; RHEUMATOID ARTHRITIS; SOLID TUMOR; THROMBOSIS;

ANIMALS; ANTIBODIES; ANTIBODIES, BLOCKING; HUMANS; RECOMBINANT FUSION PROTEINS;

EID: 54849403487     PISSN: 14714892     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.coph.2008.07.006     Document Type: Review

References (84)

1. Wu A.M., and Senter P.D. Arming antibodies: prospects and challenges for immunoconjugates. Nat Biotechnol 23 (2005) 1137-1146
2. Borrebaeck C.A., Malmborg A.C., Furebring C., Michaelsson A., Ward S., Danielsson L., and Ohlin M. Kinetic analysis of recombinant antibody-antigen interactions: relation between structural domains and antigen binding. Biotechnology (NY) 10 (1992) 697-698
3. Utsumi S., and Karush F. The subunits of purified rabbit antibody. Biochemistry 3 (1964) 1329-1338
4. Ward E.S., Gussow D., Griffiths A.D., Jones P.T., and Winter G. Binding activities of a repertoire of single immunoglobulin variable domains secreted from Escherichia coli. Nature 341 (1989) 544-546
5. Borrebaeck C.A., and Ohlin M. Antibody evolution beyond nature. Nat Biotechnol 20 (2002) 1189-1190
6. Reiter Y., Schuck P., Boyd L.F., and Plaksin D. 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 (1999) 685-698
7. Soderlind E., Vergeles M., and Borrebaeck C.A. Domain libraries: synthetic diversity for de novo design of antibody V-regions. Gene 160 (1995) 269-272
8. Holt L.J., Herring C., Jespers L.S., Woolven B.P., and Tomlinson I.M. Domain antibodies: proteins for therapy. Trends Biotechnol 21 (2003) 484-490
9. Gill D.S., and Damle N.K. Biopharmaceutical drug discovery using novel protein scaffolds. Curr Opin Biotechnol 17 (2006) 653-658. This article reviews the pros and cons and the application of novel scaffold technologies in current drug development. Major scaffolds such as single-domain antibodies, tetranectins, ankyrins, and lipocalins are discussed.
10. Hamers-Casterman C., Atarhouch T., Muyldermans S., Robinson G., Hamers C., Songa E.B., Bendahman N., and Hamers R. Naturally occurring antibodies devoid of light chains. Nature 363 (1993) 446-448
11. Arbabi Ghahroudi M., Desmyter A., Wyns L., Hamers R., and Muyldermans S. Selection and identification of single domain antibody fragments from camel heavy-chain antibodies. FEBS Lett 414 (1997) 521-526
12. Muyldermans S. Single domain camel antibodies: current status. J Biotechnol 74 (2001) 277-302
13. Harmsen M.M., and De Haard H.J. Properties, production, and applications of camelid single-domain antibody fragments. Appl Microbiol Biotechnol 77 (2007) 13-22
14. Kopsidas G., Carman R.K., Stutt E.L., Raicevic A., Roberts A.S., Siomos M.A., Dobric N., Pontes-Braz L., and Coia G. RNA mutagenesis yields highly diverse mRNA libraries for in vitro protein evolution. BMC Biotechnol 7 (2007) 18
15. Yau K.Y., Groves M.A., Li S., Sheedy C., Lee H., Tanha J., MacKenzie C.R., Jermutus L., and Hall J.C. Selection of hapten-specific single-domain antibodies from a non-immunized llama ribosome display library. J Immunol Methods 281 (2003) 161-175
16. Dooley H., Flajnik M.F., and Porter A.J. Selection and characterization of naturally occurring single-domain (IgNAR) antibody fragments from immunized sharks by phage display. Mol Immunol 40 (2003) 25-33
17. Nuttall S.D., Krishnan U.V., Doughty L., Pearson K., Ryan M.T., Hoogenraad N.J., Hattarki M., Carmichael J.A., Irving R.A., and Hudson P.J. Isolation and characterization of an IgNAR variable domain specific for the human mitochondrial translocase receptor Tom70. Eur J Biochem 270 (2003) 3543-3554
18. Shao C.Y., Secombes C.J., and Porter A.J. Rapid isolation of IgNAR variable single-domain antibody fragments from a shark synthetic library. Mol Immunol 44 (2007) 656-665
19. Conrath K., Vincke C., Stijlemans B., Schymkowitz J., Decanniere K., Wyns L., Muyldermans S., and Loris R. Antigen binding and solubility effects upon the veneering of a camel VHH in framework-2 to mimic a VH. J Mol Biol 350 (2005) 112-125
20. Kruger C., Hultberg A., van Dollenweerd C., Marcotte H., and Hammarstrom L. Passive immunization by lactobacilli expressing single-chain antibodies against Streptococcus mutans. Mol Biotechnol 31 (2005) 221-231
21. Pant N., Hultberg A., Zhao Y., Svensson L., Pan-Hammarstrom Q., Johansen K., Pouwels P.H., Ruggeri F.M., Hermans P., Frenken L., et al. Lactobacilli expressing variable domain of llama heavy-chain antibody fragments (lactobodies) confer protection against rotavirus-induced diarrhea. J Infect Dis 194 (2006) 1580-1588
22. van der Vaart J.M., Pant N., Wolvers D., Bezemer S., Hermans P.W., Bellamy K., Sarker S.A., van der Logt C.P., Svensson L., Verrips C.T., et al. Reduction in morbidity of rotavirus induced diarrhoea in mice by yeast produced monovalent llama-derived antibody fragments. Vaccine 24 (2006) 4130-4137
23. Roovers R.C., Laeremans T., Huang L., De Taeye S., Verkleij A.J., Revets H., de Haard H.J., and van Bergen en Henegouwen P.M. Efficient inhibition of EGFR signaling and of tumour growth by antagonistic anti-EFGR nanobodies. Cancer Immunol Immunother 56 (2007) 303-317
24. Dumoulin M., Last A.M., Desmyter A., Decanniere K., Canet D., Larsson G., Spencer A., Archer D.B., Sasse J., Muyldermans S., et al. A camelid antibody fragment inhibits the formation of amyloid fibrils by human lysozyme. Nature 424 (2003) 783-788
25. Verheesen P., de Kluijver A., van Koningsbruggen S., de Brij M., de Haard H.J., van Ommen G.J., van der Maarel S.M., and Verrips C.T. Prevention of oculopharyngeal muscular dystrophy-associated aggregation of nuclear polyA-binding protein with a single-domain intracellular antibody. Hum Mol Genet 15 (2006) 105-111
26. Els Conrath K., Lauwereys M., Wyns L., and Muyldermans S. Camel single-domain antibodies as modular building units in bispecific and bivalent antibody constructs. J Biol Chem 276 (2001) 7346-7350
27. Simmons D.P., Abregu F.A., Krishnan U.V., Proll D.F., Streltsov V.A., Doughty L., Hattarki M.K., and Nuttall S.D. Dimerisation strategies for shark IgNAR single domain antibody fragments. J Immunol Methods 315 (2006) 171-184
28. Holt L.J., Basran A., Jones K., Chorlton J., Jespers L.S., Brewis N.D., and Tomlinson I.M. Anti-serum albumin domain antibodies for extending the half-lives of short lived drugs. Protein Eng Des Sel 21 (2008) 283-288. Using single-domain antibodies against serum albumin, the authors show that bispecific constructs with these sdAbs can match the half-life of serum albumin itself. This demonstrates the perspective to prolong the half-life of different drugs to improve the dosing regimen and/or clinical effect.
29. Coppieters K., Dreier T., Silence K., de Haard H., Lauwereys M., Casteels P., Beirnaert E., Jonckheere H., Van de Wiele C., Staelens L., et al. 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 (2006) 1856-1866. This paper explored the preclinical application of camelid anti-TNF VHH proteins as TNF antagonists in a mouse model of rheumatoid arthritis (RA). The antagonistic potency of the anti-human TNF VHH proteins exceeded even that of the anti-TNF antibodies Infliximab and Adalimumab that are used clinically in RA.
30. Zhang J., Tanha J., Hirama T., Khieu N.H., To R., Tong-Sevinc H., Stone E., Brisson J.R., and MacKenzie C.R. Pentamerization of single-domain antibodies from phage libraries: a novel strategy for the rapid generation of high-avidity antibody reagents. J Mol Biol 335 (2004) 49-56
31. Stewart C.S., MacKenzie C.R., and Hall J.C. Isolation, characterization and pentamerization of alpha-cobrotoxin specific single-domain antibodies from a naive phage display library: preliminary findings for antivenom development. Toxicon 49 (2007) 699-709
32. Stone E., Hirama T., Tanha J., Tong-Sevinc H., Li S., MacKenzie C.R., and Zhang J. The assembly of single domain antibodies into bispecific decavalent molecules. J Immunol Methods 318 (2007) 88-94
33. Behar G., Siberil S., Groulet A., Chames P., Pugniere M., Boix C., Sautes-Fridman C., Teillaud J.L., and Baty D. Isolation and characterization of anti-FcgammaRIII (CD16) llama single-domain antibodies that activate natural killer cells. Protein Eng Des Sel 21 (2008) 1-10. In this work, the authors report the isolation of anti-FcγRIII single-domain antibodies from an immunized llama. With the engagement of two single-domain antibodies to FcγRIII, they show a strong induction of IL-2 and IFN-γ production. These sdAbs can be used in bispecific constructs for the recruitment of FcγRIII killer cells to destroy the target cells of the second sdAb.
34. Harmsen M.M., Van Solt C.B., Fijten H.P., and Van Setten M.C. Prolonged in vivo residence times of llama single-domain antibody fragments in pigs by binding to porcine immunoglobulins. Vaccine 23 (2005) 4926-4934
35. Muruganandam A., Tanha J., Narang S., and Stanimirovic D. Selection of phage-displayed llama single-domain antibodies that transmigrate across human blood-brain barrier endothelium. FASEB J 16 (2002) 240-242
36. Abulrob A., Sprong H., Van Bergen en Henegouwen P., and Stanimirovic D. The blood-brain barrier transmigrating single domain antibody: mechanisms of transport and antigenic epitopes in human brain endothelial cells. J Neurochem 95 (2005) 1201-1214
37. Miao Q.F., Liu X.Y., Shang B.Y., Ouyang Z.G., and Zhen Y.S. An enediyne-energized single-domain antibody-containing fusion protein shows potent antitumor activity. Anticancer Drugs 18 (2007) 127-137. The tumor-targeting potency of a type IV collagenase specific sdAb was demonstrated after its linkage to labile enediyne chromophore (AE) and a noncovalent bound apoprotein (LDP). The VH-LDP-AE construct showed extremely potent cytotoxicity to cancer cells and marked antiangiogenic activity in vitro. This construct would be a promising candidate for cancer-targeting therapy.
38. Cortez-Retamozo V., Backmann N., Senter P.D., Wernery U., De Baetselier P., Muyldermans S., and Revets H. Efficient cancer therapy with a nanobody-based conjugate. Cancer Res 64 (2004) 2853-2857
39. Szynol A., de Haard J.J., Veerman E.C., de Soet J.J., and van Nieuw Amerongen A.V. Design of a peptibody consisting of the antimicrobial peptide dhvar5 and a llama variable heavy-chain antibody fragment. Chem Biol Drug Des 67 (2006) 425-431
40. Baral T.N., Magez S., Stijlemans B., Conrath K., Vanhollebeke B., Pays E., Muyldermans S., and De Baetselier P. Experimental therapy of African trypanosomiasis with a nanobody-conjugated human trypanolytic factor. Nat Med 12 (2006) 580-584
41. Suter M., Blaser K., Aeby P., and Crameri R. Rabbit single domain antibodies specific to protein C expressed in prokaryotes. Immunol Lett 33 (1992) 53-59
42. Kieber-Emmons T., von Feldt J.M., Godillot A.P., McCallus D., Srikantan V., Weiner D.B., and Williams W.V. Isolated VH4 heavy chain variable regions bind DNA characterization of a recombinant antibody heavy chain library derived from patient(s) with active SLE. Lupus 3 (1994) 379-392
43. Davies J., and Riechmann L. Antibody VH domains as small recognition units. Biotechnology (NY) 13 (1995) 475-479
44. Martin F., Volpari C., Steinkuhler C., Dimasi N., Brunetti M., Biasiol G., Altamura S., Cortese R., De Francesco R., and Sollazzo M. Affinity selection of a camelized V(H) domain antibody inhibitor of hepatitis C virus NS3 protease. Protein Eng 10 (1997) 607-614
45. Lauwereys M., Arbabi Ghahroudi M., Desmyter A., Kinne J., Holzer W., De Genst E., Wyns L., and Muyldermans S. Potent enzyme inhibitors derived from dromedary heavy-chain antibodies. EMBO J 17 (1998) 3512-3520
46. van den Beucken T., van Neer N., Sablon E., Desmet J., Celis L., Hoogenboom H.R., and Hufton S.E. Building novel binding ligands to B7.1 and B7. 2 based on human antibody single variable light chain domains. J Mol Biol 310 (2001) 591-601
47. Tanha J., Dubuc G., Hirama T., Narang S.A., and MacKenzie C.R. Selection by phage display of llama conventional V(H) fragments with heavy chain antibody V(H)H properties. J Immunol Methods 263 (2002) 97-109
48. Desmyter A., Spinelli S., Payan F., Lauwereys M., Wyns L., Muyldermans S., and Cambillau C. Three camelid VHH domains in complex with porcine pancreatic alpha-amylase. Inhibition and versatility of binding topology. J Biol Chem 277 (2002) 23645-23650
49. Tanaka T., Lobato M.N., and Rabbitts T.H. Single domain intracellular antibodies: a minimal fragment for direct in vivo selection of antigen-specific intrabodies. J Mol Biol 331 (2003) 1109-1120
50. Verheesen P., ten Haaft M.R., Lindner N., Verrips C.T., and de Haard J.J. Beneficial properties of single-domain antibody fragments for application in immunoaffinity purification and immuno-perfusion chromatography. Biochim Biophys Acta 1624 (2003) 21-28
51. Dekker S., Toussaint W., Panayotou G., de Wit T., Visser P., Grosveld F., and Drabek D. Intracellularly expressed single-domain antibody against p15 matrix protein prevents the production of porcine retroviruses. J Virol 77 (2003) 12132-12139
52. van Koningsbruggen S., de Haard H., de Kievit P., Dirks R.W., van Remoortere A., Groot A.J., van Engelen B.G., den Dunnen J.T., Verrips C.T., Frants R.R., et al. Llama-derived phage display antibodies in the dissection of the human disease oculopharyngeal muscular dystrophy. J Immunol Methods 279 (2003) 149-161
53. Lah J., Marianovsky I., Glaser G., Engelberg-Kulka H., Kinne J., Wyns L., and Loris R. Recognition of the intrinsically flexible addiction antidote MazE by a dromedary single domain antibody fragment. Structure, thermodynamics of binding, stability, and influence on interactions with DNA. J Biol Chem 278 (2003) 14101-14111
54. Jobling S.A., Jarman C., Teh M.M., Holmberg N., Blake C., and Verhoeyen M.E. Immunomodulation of enzyme function in plants by single-domain antibody fragments. Nat Biotechnol 21 (2003) 77-80
55. Huang M., Zhang B., Wang J., Mei F., and Hou L. Development of single-domain recombinant antibodies to reverse transcriptase domain of human hTERT. Hybrid Hybridomics 23 (2004) 244-249
56. Rahbarizadeh F., Rasaee M.J., Forouzandeh Moghadam M., Allameh A.A., and Sadroddiny E. Production of novel recombinant single-domain antibodies against tandem repeat region of MUC1 mucin. Hybrid Hybridomics 23 (2004) 151-159
57. Omidfar K., Rasaee M.J., Modjtahedi H., Forouzandeh M., Taghikhani M., Bakhtiari A., Paknejad M., and Kashanian S. Production and characterization of a new antibody specific for the mutant EGF receptor, EGFRvIII, in Camelus bactrianus. Tumour Biol 25 (2004) 179-187
58. Saerens D., Kinne J., Bosmans E., Wernery U., Muyldermans S., and Conrath K. Single domain antibodies derived from dromedary lymph node and peripheral blood lymphocytes sensing conformational variants of prostate-specific antigen. J Biol Chem 279 (2004) 51965-51972
59. Stijlemans B., Conrath K., Cortez-Retamozo V., Van Xong H., Wyns L., Senter P., Revets H., De Baetselier P., Muyldermans S., and Magez S. Efficient targeting of conserved cryptic epitopes of infectious agents by single domain antibodies. African trypanosomes as paradigm. J Biol Chem 279 (2004) 1256-1261
60. Colby D.W., Garg P., Holden T., Chao G., Webster J.M., Messer A., Ingram V.M., and Wittrup K.D. Development of a human light chain variable domain (V(L)) intracellular antibody specific for the amino terminus of huntingtin via yeast surface display. J Mol Biol 342 (2004) 901-912
61. Paz K., Brennan L.A., Iacolina M., Doody J., Hadari Y.R., and Zhu Z. Human single-domain neutralizing intrabodies directed against Etk kinase: a novel approach to impair cellular transformation. Mol Cancer Ther 4 (2005) 1801-1809
62. De Haard H.J., Bezemer S., Ledeboer A.M., Muller W.H., Boender P.J., Moineau S., Coppelmans M.C., Verkleij A.J., Frenken L.G., and Verrips C.T. Llama antibodies against a lactococcal protein located at the tip of the phage tail prevent phage infection. J Bacteriol 187 (2005) 4531-4541
63. Zarebski L.M., Urrutia M., and Goldbaum F.A. Llama single domain antibodies as a tool for molecular mimicry. J Mol Biol 349 (2005) 814-824
64. Dolk E., van der Vaart M., Lutje Hulsik D., Vriend G., de Haard H., Spinelli S., Cambillau C., Frenken L., and Verrips T. Isolation of llama antibody fragments for prevention of dandruff by phage display in shampoo. Appl Environ Microbiol 71 (2005) 442-450
65. Shuntao W., Jiannan F., Jianwei G., Leiming G., Yan L., Yingxun S., Weisong Q., Meiru H., Gencheng H., and Beifen S. A novel designed single domain antibody on 3-D structure of ricin A chain remarkably blocked ricin-induced cytotoxicity. Mol Immunol 43 (2006) 1912-1919
66. Kim Y.R., Kim J.S., Lee S.H., Lee W.R., Sohn J.N., Chung Y.C., Shim H.K., Lee S.C., Kwon M.H., and Kim Y.S. Heavy and light chain variable single domains of an anti-DNA binding antibody hydrolyze both double- and single-stranded DNAs without sequence specificity. J Biol Chem 281 (2006) 15287-15295
67. Goldman E.R., Anderson G.P., Liu J.L., Delehanty J.B., Sherwood L.J., Osborn L.E., Cummins L.B., and Hayhurst A. Facile generation of heat-stable antiviral and antitoxin single domain antibodies from a semisynthetic llama library. Anal Chem 78 (2006) 8245-8255
68. Mai K.T., Perkins D.G., Zhang J., and Mackenzie C.R. ES1, a new lung carcinoma antibody - an immunohistochemical study. Histopathology 49 (2006) 515-522
69. Gueorguieva D., Li S., Walsh N., Mukerji A., Tanha J., and Pandey S. Identification of single-domain, Bax-specific intrabodies that confer resistance to mammalian cells against oxidative-stress-induced apoptosis. FASEB J 20 (2006) 2636-2638
70. Groot A.J., Verheesen P., Westerlaken E.J., Gort E.H., van der Groep P., Bovenschen N., van der Wall E., van Diest P.J., and Shvarts A. Identification by phage display of single-domain antibody fragments specific for the ODD domain in hypoxia-inducible factor 1alpha. Lab Invest 86 (2006) 345-356
71. Harmsen M.M., van Solt C.B., van Zijderveld-van Bemmel A.M., Niewold T.A., and van Zijderveld F.G. Selection and optimization of proteolytically stable llama single-domain antibody fragments for oral immunotherapy. Appl Microbiol Biotechnol 72 (2006) 544-551
72. Ladenson R.C., Crimmins D.L., Landt Y., and Ladenson J.H. Isolation and characterization of a thermally stable recombinant anti-caffeine heavy-chain antibody fragment. Anal Chem 78 (2006) 4501-4508
73. Anderson G.P., Matney R., Liu J.L., Hayhurst A., and Goldman E.R. Multiplexed fluid array screening of phage displayed anti-ricin single domain antibodies for rapid assessment of specificity. Biotechniques 43 (2007) 806-811
74. Sherwood L.J., Osborn L.E., Carrion Jr. R., Patterson J.L., and Hayhurst A. Rapid assembly of sensitive antigen-capture assays for Marburg virus, using in vitro selection of llama single-domain antibodies, at biosafety level 4. J Infect Dis 196 Suppl 2 (2007) S213-219
75. Koch-Nolte F., Reyelt J., Schossow B., Schwarz N., Scheuplein F., Rothenburg S., Haag F., Alzogaray V., Cauerhff A., and Goldbaum F.A. Single domain antibodies from llama effectively and specifically block T cell ecto-ADP-ribosyltransferase ART2.2 in vivo. FASEB J 21 (2007) 3490-3498
76. Ismaili A., Jalali-Javaran M., Rasaee M.J., Rahbarizadeh F., Forouzandeh-Moghadam M., and Memari H.R. Production and characterization of anti-(mucin MUC1) single-domain antibody in tobacco (Nicotiana tabacum cultivar Xanthi). Biotechnol Appl Biochem 47 (2007) 11-19
77. Alvarez-Rueda N., Behar G., Ferre V., Pugniere M., Roquet F., Gastinel L., Jacquot C., Aubry J., Baty D., Barbet J., et al. Generation of llama single-domain antibodies against methotrexate, a prototypical hapten. Mol Immunol 44 (2007) 1680-1690
78. Cossins A.J., Harrison S., Popplewell A.G., and Gore M.G. Recombinant production of a VL single domain antibody in Escherichia coli and analysis of its interaction with peptostreptococcal protein L. Protein Expr Purif 51 (2007) 253-259
79. Liu J.L., Anderson G.P., and Goldman E.R. Isolation of anti-toxin single domain antibodies from a semi-synthetic spiny dogfish shark display library. BMC Biotechnol 7 (2007) 78
80. Henderson K.A., Streltsov V.A., Coley A.M., Dolezal O., Hudson P.J., Batchelor A.H., Gupta A., Bai T., Murphy V.J., Anders R.F., et al. Structure of an IgNAR-AMA1 complex: targeting a conserved hydrophobic cleft broadens malarial strain recognition. Structure 15 (2007) 1452-1466
81. Liu J.L., Anderson G.P., Delehanty J.B., Baumann R., Hayhurst A., and Goldman E.R. Selection of cholera toxin specific IgNAR single-domain antibodies from a naive shark library. Mol Immunol 44 (2007) 1775-1783
82. Inoue A., Sawata S.Y., Taira K., and Wadhwa R. Loss-of-function screening by randomized intracellular antibodies: identification of hnRNP-K as a potential target for metastasis. Proc Natl Acad Sci U S A 104 (2007) 8983-8988
83. Saerens D., Stijlemans B., Baral T.N., Nguyen Thi G.T., Wernery U., Magez S., De Baetselier P., Muyldermans S., and Conrath K. Parallel selection of multiple anti-infectome Nanobodies without access to purified antigens. J Immunol Methods 329 (2008) 138-150
84. Hmila I., Ben Abderrazek R., Saerens D., Benlasfar Z., Conrath K., El Ayeb M., Muyldermans S., and Bouhaouala-Zahar B. VHH, bivalent domains and chimeric heavy-chain-only antibodies with high neutralizing efficacy for scorpion toxin AahI. Mol. Immunol. 45 (2008) 3847-3856