The development of nanobodies for therapeutic applications

Current Opinion in Investigational Drugs

Volumn 10, Issue 11, 2009, Pages 1212-1224

  Van Bockstaele, F ^a   Holz, J B ^a   Revets, H ^a  

^a ABLYNX NV   (Belgium)

Author keywords

Heavy chain antibody; Nanobody; Single domain antibody; Therapeutic; V HH

Indexed keywords

ALX 0081; ALX 0141; ALX 0681; ANTICOAGULANT AGENT; EPITOPE; FC RECEPTOR; IMMUNOGLOBULIN; NANOBODY; PLACEBO; RECOMBINANT ANTIBODY; UNCLASSIFIED DRUG;

ALZHEIMER DISEASE; ANTIBODY STRUCTURE; BLOOD BRAIN BARRIER; BONE METASTASIS; CANCER RESEARCH; CLINICAL TRIAL; CONFORMATIONAL TRANSITION; DRUG DELIVERY SYSTEM; DRUG DESIGN; DRUG DISTRIBUTION; DRUG MANUFACTURE; DRUG RESEARCH; DRUG SAFETY; DRUG SOLUBILITY; DRUG STABILITY; DRUG STRUCTURE; DRUG TARGETING; DRUG TOLERABILITY; EFFECTOR CELL; HEAVY CHAIN; HUMAN; IMMUNE SYSTEM; MEDICAL TECHNOLOGY; MOLECULAR WEIGHT; NONHUMAN; OSTEOPOROSIS; REVIEW; RHEUMATOID ARTHRITIS; THROMBOSIS;

ANIMALS; ANTIBODIES; CAMELIDS, NEW WORLD; CLINICAL TRIALS, PHASE I AS TOPIC; DRUG DESIGN; FIBRINOLYTIC AGENTS; HUMANS; IMMUNOGLOBULIN FRAGMENTS; IMMUNOGLOBULIN HEAVY CHAINS;

EID: 70449429345     PISSN: 14724472     EISSN: None     Source Type: Journal    
DOI: None     Document Type: Review

References (151)

1. Kohler G, Milstein C: Continuous cultures of fused cells secreting antibody of predefned specifcity. Nature (1975) 256(5517):495-497.
2. Khazaeli MB, Conry RM, LoBuglio AF: Human immune response to monoclonal antibodies. J Immunother Emphasis Tumor Immunol (1994) 15(1):42-52.
3. Boulianne GL, Hozumi N, Shulman MJ: Production of functional chimaeric mouse/human antibody. Nature (1984) 312(5995): 643-646.
4. Padlan EA: A possible procedure for reducing the immunogenicity of antibody variable domains while preserving their ligand-binding properties. Mol Immunol (1991) 28(4-5): 489-498.
5. Riechmann L, Clark M, Waldmann H, Winter G: Reshaping human antibodies for therapy. Nature (1988) 332(6162):323-327.
6. Green LL, Hardy MC, Maynard-Currie CE, Tsuda H, Louie DM, Mendez MJ, Abderrahim H, Noguchi M, Smith DH, Zeng Y, David NE et al: Antigen-specifc human monoclonal antibodies from mice engineered with human Ig heavy and light chain YACs. Nat Genet (1994) 7(1):13-21.
7. Huls GA, Heijnen IA, Cuomo ME, Koningsberger JC, Wiegman L, Boel E, van der Vuurst de Vries AR, Loyson SA, Helfrich W, van Berge Henegouwen GP, van Meijer M et al: A recombinant, fully human monoclonal antibody with antitumor activity constructed from phage-displayed antibody fragments. Nat Biotechnol (1999) 17(3):276-281.
8. Reichert JM, Rosensweig CJ, Faden LB, Dewitz MC: Monoclonal antibody successes in the clinic. Nat Biotechnol (2005) 23(9):1073-1078.
9. Better M, Chang CP, Robinson RR, Horwitz AH: Escherichia coli secretion of an active chimeric antibody fragment. Science (1988) 240(4855):1041-1043.
10. Skerra A, Pluckthun A: Assembly of a functional immunoglobulin Fv fragment in Escherichia coli. Science (1988) 240(4855):1038-1041.
11. Bird RE, Hardman KD, Jacobson JW, Johnson S, Kaufman BM, Lee SM, Lee T, Pope SH, Riordan GS, Whitlow M: Single-chain antigen-binding proteins. Science (1988) 242(4877):423-426.
12. Ward ES, Gussow D, Griffths AD, Jones PT, Winter G: Binding activities of a repertoire of single immunoglobulin variable domains secreted from Escherichia coli. Nature (1989) 341(6242):544-546. (Pubitemid 19252529)
13. Borrebaeck CA, Malmborg AC, Furebring C, Michaelsson A, Ward S, Danielsson L, Ohlin M: Kinetic analysis of recombinant antibody-antigen interactions: Relation between structural domains and antigen binding. Biotechnology (NY) (1992) 10(6):697-698.
14. Hamers-Casterman C, Atarhouch T, Muyldermans S, Robinson G, Hamers C, Songa EB, Bendahman N, Hamers R: Naturally occurring antibodies devoid of light chains. Nature (1993) 363(6428):446-448. •• Describes the discovery of heavy-chain antibodies in camelids.
15. Nguyen VK, Hamers R, Wyns L, Muyldermans S: Loss of splice consensus signal is responsible for the removal of the entire CH1 domain of the functional camel IgG2A heavy-chain antibodies. Mol Immunol (1999) 36(8):515-524.
16. Woolven BP, Frenken LG, van der Logt P, Nicholls PJ: The structure of the llama heavy chain constant genes reveals a mechanism for heavy-chain antibody formation. Immunogenetics (1999) 50(1-2):98-101.
17. Padlan EA: Anatomy of the antibody molecule. Mol Immunol (1994) 31(3):169-217.
18. Muyldermans S: Single domain camel antibodies: Current status. J Biotechnol (2001) 74(4):277-302.
19. Nguyen VK, Hamers R, Wyns L, Muyldermans S: Camel heavy-chain antibodies: Diverse germline VHH and specifc mechanisms enlarge the antigen-binding repertoire. Embo J (2000) 19(5):921-930.
20. Muyldermans S, Atarhouch T, Saldanha J, Barbosa JA, Hamers R: Sequence and structure of VH domain from naturally occurring camel heavy chain immunoglobulins lacking light chains. Protein Eng (1994) 7(9):1129-1135. (Pubitemid 24282500)
21. Vu KB, Ghahroudi MA, Wyns L, Muyldermans S: Comparison of llama VH sequences from conventional and heavy chain antibodies. Mol Immunol (1997) 34(16-17):1121-1131.
22. Nguyen VK, Desmyter A, Muyldermans S: Functional heavy-chain antibodies in Camelidae. In: Advances in immunology. Dixon F (Ed), Academic Press, London, UK (2001) 79:261-296.
23. Desmyter A, Transue TR, Ghahroudi MA, Thi MH, Poortmans F, Hamers R, Muyldermans S, Wyns L: Crystal structure of a camel single-domain VH antibody fragment in complex with lysozyme. Nat Struct Biol (1996) 3(9):803-811.
24. Harmsen MM, Ruuls RC, Nijman IJ, Niewold TA, Frenken LG, de Geus B: Llama heavy-chain V regions consist of at least four distinct subfamilies revealing novel sequence features. Mol Immunol (2000) 37(10):579-590.
25. Decanniere K, Desmyter A, Lauwereys M, Ghahroudi MA, Muyldermans S, Wyns L: A single-domain antibody fragment in complex with RNase A: Non-canonical loop structures and nanomolar affnity using two CDR loops. Structure (1999) 7(4):361-370.
26. Spinelli S, Frenken L, Bourgeois D, de Ron L, Bos W, Verrips T, Anguille C, Cambillau C, Tegoni M: The crystal structure of a llama heavy chain variable domain. Nat Struct Biol (1996) 3(9):752-757.
27. Spinelli S, Frenken LG, Hermans P, Verrips T, Brown K, Tegoni M, Cambillau C: Camelid heavy-chain variable domains provide effcient combining sites to haptens. Biochemistry (2000) 39(6):1217-1222.
28. Arbabi Ghahroudi M, Desmyter A, Wyns L, Hamers R, Muyldermans S: Selection and identifcation of single domain antibody fragments from camel heavy-chain antibodies. FEBS Lett (1997) 414(3):521-526.
29. Lauwereys M, Arbabi Ghahroudi M, Desmyter A, Kinne J, Holzer W, De Genst E, Wyns L, Muyldermans S: Potent enzyme inhibitors derived from dromedary heavy-chain antibodies. Embo J (1998) 17(13):3512-3520.
30. Omidfar K, Rasaee MJ, Modjtahedi H, Forouzandeh M, Taghikhani M, Bakhtiari A, Paknejad M, Kashanian S: Production and characterization of a new antibody specifc for the mutant EGF receptor, EGFRvIII, in Camelus bactrianus. Tumor Biol (2004) 25(4):179-187.
31. Rahbarizadeh F, Rasaee MJ, Forouzandeh Moghadam M, Allameh AA, Sadroddiny E: Production of novel recombinant single-domain antibodies against tandem repeat region of MUC1 mucin. Hybrid Hybridomics (2004) 23(3):151-159.
32. van der Linden R, de Geus B, Stok W, Bos W, van Wassenaar D, Verrips T, Frenken L: Induction of immune responses and molecular cloning of the heavy chain antibody repertoire of Lama glama. J Immunol Methods (2000) 240(1-2):185-195.
33. Maass DR, Sepulveda J, Pernthaner A, Shoemaker CB: Alpaca (Lama pacos) as a convenient source of recombinant camelid heavy chain antibodies (VHHs). J Immunol Methods (2007) 324(1-2):13-25.
34. Tanha J, Dubuc G, Hirama T, Narang SA, MacKenzie CR: Selection by phage display of llama conventional VH fragments with heavy chain antibody VHH properties. J Immunol Methods (2002) 263(1-2):97-109.
35. Verheesen P, de Kluijver A, van Koningsbruggen S, de Brij M, de Haard HJ, van Ommen GJ, van der Maarel SM, Verrips CT: Prevention of oculopharyngeal muscular dystrophy-associated aggregation of nuclear polyA-binding protein with a single-domain intracellular antibody. Hum Mol Genet (2006) 15(1):105-111.
36. Verheesen P, Roussis A, de Haard HJ, Groot AJ, Stam JC, den Dunnen JT, Frants RR, Verkleij AJ, Theo Verrips C, van der Maarel SM: Reliable and controllable antibody fragment selections from camelid non-immune libraries for target validation. Biochim Biophys Acta (2006) 1764(8):1307-1319.
37. Yau KY, Groves MA, Li S, Sheedy C, Lee H, Tanha J, MacKenzie CR, Jermutus L, Hall JC: Selection of hapten-specifc single-domain antibodies from a non-immunized llama ribosome display library. J Immunol Methods (2003) 281(1-2):161-175.
38. Davies J, Riechmann L: Single antibody domains as small recognition units: Design and in vitro antigen selection of camelized, human VH domains with improved protein stability. Protein Eng (1996) 9(6):531-537.
39. Goldman ER, Anderson GP, Liu JL, Delehanty JB, Sherwood LJ, Osborn LE, Cummins LB, Hayhurst A: Facile generation of heat-stable antiviral and antitoxin single domain antibodies from a semisynthetic llama library. Anal Chem (2006) 78(24):8245-8255.
40. Martin F, Volpari C, Steinkuhler C, Dimasi N, Brunetti M, Biasiol G, Altamura S, Cortese R, De Francesco R, Sollazzo M: Affnity selection of a camelized VH domain antibody inhibitor of hepatitis C virus NS3 protease. Protein Eng (1997) 10(5): 607-614.
41. Tanha J, Xu P, Chen Z, Ni F, Kaplan H, Narang SA, MacKenzie CR: Optimal design features of camelized human single-domain antibody libraries. J Biol Chem (2001) 276(27):24774-24780.
42. Research: Ablynx NV, Ghent, Zwijnaarde, Belgium (2008). www. ablynx.com/research/research.htm
43. Vincke C, Loris R, Saerens D, Martinez-Rodriguez S, Muyldermans S, Conrath K: General strategy to humanize a camelid single-domain antibody and identifcation of a universal humanized Nanobody scaffold. J Biol Chem (2008) 284(5):3273-3284.
44. Davies J, Riechmann L: 'Camelising' human antibody fragments: NMR studies on VH domains. FEBS Lett (1994) 339(3):285-290.
45. Davies J, Riechmann L: Antibody VH domains as small recognition units. Biotechnology (NY) (1995) 13(5):475-479.
46. Frenken LG, van der Linden RH, Hermans PW, Bos JW, Ruuls RC, de Geus B, Verrips CT: Isolation of antigen specifc llama VHH antibody fragments and their high level secretion by Saccharomyces cerevisiae. J Biotechnol (2000) 78(1):11-21.
47. Thomassen YE, Meijer W, Sierkstra L, Verrips CT: Large-scale production of VHH antibody fragments by Saccharomyces cerevisiae. Enzym Microb Tech (2002) 30(3):273-278.
48. van der Vaart JM: Expression of VHH antibody fragments in Saccharomyces cerevisiae. In: Methods in molecular biology. O'Brien PM, Aitken R (Eds), Humana Press Inc, Totowa, NJ, USA (2002) 178:359-366.
49. Bazl MR, Rasaee MJ, Foruzandeh M, Rahimpour A, Kiani J, Rahbarizadeh F, Alirezapour B, Mohammadi M: Production of chimeric recombinant single domain antibody-green fuorescent fusion protein in Chinese hamster ovary cells. Hybridoma (Larchmt) (2007) 26(1):1-9.
50. Omidfar K, Rasaee MJ, Kashanian S, Paknejad M, Bathaie Z: Studies of thermostability in Camelus bactrianus (Bactrian camel) single-domain antibody specifc for the mutant epidermal-growth-factor receptor expressed by Pichia. Biotechnol Appl Biochem (2007) 46(1):41-49.
51. Rahbarizadeh F, Rasaee MJ, Forouzandeh M, Allameh AA: Over expression of anti-MUC1 single-domain antibody fragments in the yeast Pichia pastoris. Mol Immunol (2006) 43(5):426-435. (Pubitemid 41740431)
52. Joosten V, Gouka RJ, van den Hondel CA, Verrips CT, Lokman BC: Expression and production of llama variable heavy-chain antibody fragments (VHHs) by Aspergillus awamori. Appl Microbiol Biotechnol (2005) 66(4):384-392.
53. Joosten V, Roelofs MS, van den Dries N, Goosen T, Verrips CT, van den Hondel CA, Lokman BC: Production of bifunctional proteins by Aspergillus awamori: Llama variable heavy chain antibody fragment (VHH) R9 coupled to Arthromyces ramosus peroxidase (ARP). J Biotechnol (2005) 120(4):347-359.
54. Ismaili A, Jalali-Javaran M, Rasaee MJ, Rahbarizadeh F, Forouzandeh-Moghadam M, Memari HR: Production and characterization of anti-(mucin MUC1) single-domain antibody in tobacco (Nicotiana tabacum cultivar Xanthi). Biotechnol Appl Biochem (2007) 47(1):11-19.
55. Jobling SA, Jarman C, Teh MM, Holmberg N, Blake C, Verhoeyen ME: Immunomodulation of enzyme function in plants by single-domain antibody fragments. Nat Biotechnol (2003) 21(1):77-80.
56. Rajabi-Memari H, Jalali-Javaran M, Rasaee MJ, Rahbarizadeh F, Forouzandeh-Moghadam M, Esmaili A: Expression and characterization of a recombinant single-domain monoclonal antibody against MUC1 mucin in tobacco plants. Hybridoma (Larchmt) (2006) 25(4):209-215.
57. Winichayakul S, Pernthaner A, Scott R, Vlaming R, Roberts N: Head-to-tail fusions of camelid antibodies can be expressed in planta and bind in rumen fuid. Biotechnol Appl Biochem (2009) 53(Pt 2):111-122.
58. Harmsen MM, De Haard HJ: Properties, production, and applications of camelid single-domain antibody fragments. Appl Microbiol Biotechnol (2007) 77(1):13-22.
59. Revets H, De Baetselier P, Muyldermans S: Nanobodies as novel agents for cancer therapy. Expert Opin Biol Ther (2005) 5(1): 111-124.
60. Cortez-Retamozo V, Lauwereys M, Hassanzadeh Gh G, Gobert M, Conrath K, Muyldermans S, De Baetselier P, Revets H: Effcient tumor targeting by single-domain antibody fragments of camels. Int J Cancer (2002) 98(3):456-462.
61. Cortez-Retamozo V, Backmann N, Senter PD, Wernery U, De Baetselier P, Muyldermans S, Revets H: Effcient cancer therapy with a nanobody-based conjugate. Cancer Res (2004) 64(8):2853-2857.
62. Gainkam LO, Huang L, Caveliers V, Keyaerts M, Hernot S, Vaneycken I, Vanhove C, Revets H, De Baetselier P, Lahoutte T: Comparison of the biodistribution and tumor targeting of two 99mTc-labeled anti-EGFR nanobodies in mice, using pinhole SPECT/micro-CT. J Nucl Med (2008) 49(5):788-795.
63. Huang L, Gainkam LO, Caveliers V, Vanhove C, Keyaerts M, De Baetselier P, Bossuyt A, Revets H, Lahoutte T: SPECT imaging with 99mTc-labeled EGFR-specifc nanobody for in vivo monitoring of EGFR expression. Mol Imaging Biol (2008) 10(3):167-175.
64. Rothbauer U, Zolghadr K, Tillib S, Nowak D, Schermelleh L, Gahl A, Backmann N, Conrath K, Muyldermans S, Cardoso MC, Leonhardt H: Targeting and tracing antigens in live cells with fuorescent nanobodies. Nat Methods (2006) 3(11):887-889.
65. Harmsen MM, van Solt CB, Fijten HP, van Keulen L, Rosalia RA, Weerdmeester K, Cornelissen AH, De Bruin MG, Eble PL, Dekker A: Passive immunization of guinea pigs with llama single-domain antibody fragments against foot-and-mouth disease. Vet Microbiol (2007) 120(3-4):193-206.
66. Harmsen MM, Fijten HP, Dekker A, Eble PL: Passive immunization of pigs with bispecifc llama single-domain antibody fragments against foot-and-mouth disease and porcine immunoglobulin. Vet Microbiol (2008) 132(1-2):56-64.
67. Harmsen MM, Fijten HP, Engel B, Dekker A, Eble PL: Passive immunization with llama single-domain antibody fragments reduces foot-and-mouth disease transmission between pigs. Vaccine (2009) 27(13):1904-1911.
68. Hoogenboom HR: Mix and match: Building manifold binding sites. Nat Biotechnol (1997) 15(2):125-126.
69. Tijink BM, Laeremans T, Budde M, Stigter-van Walsum M, Dreier T, de Haard HJ, Leemans CR, van Dongen GA: Improved tumor targeting of anti-epidermal growth factor receptor nanobodies through albumin binding: Taking advantage of modular nanobody technology. Mol Cancer Ther (2008) 7(8): 2288-2297.
70. Roovers RC, Laeremans T, Huang L, De Taeye S, Verkleij AJ, Revets H, de Haard HJ, van Bergen en Henegouwen PM: Effcient inhibition of EGFR signaling and of tumour growth by antagonistic anti-EFGR nanobodies. Cancer Immunol Immunother (2007) 56(3):303-317.
71. Alvarez-Rueda N, Behar G, Ferre V, Pugniere M, Roquet F, Gastinel L, Jacquot C, Aubry J, Baty D, Barbet J, Birkle S: Generation of llama single-domain antibodies against methotrexate, a prototypical hapten. Mol Immunol (2007) 44(7):1680-1690.
72. Doyle PJ, Arbabi-Ghahroudi M, Gaudette N, Furzer G, Savard ME, Gleddie S, McLean MD, Mackenzie CR, Hall JC: Cloning, expression, and characterization of a single-domain antibody fragment with affnity for 15-acetyl-deoxynivalenol. Mol Immunol (2008) 45(14):3703-3713.
73. Spinelli S, Tegoni M, Frenken L, van Vliet C, Cambillau C: Lateral recognition of a dye hapten by a llama VHH domain. J Mol Biol (2001) 311(1):123-129.
74. Stijlemans B, Conrath K, Cortez-Retamozo V, Van Xong H, Wyns L, Senter P, Revets H, De Baetselier P, Muyldermans S, Magez S: Effcient targeting of conserved cryptic epitopes of infectious agents by single domain antibodies. African trypanosomes as paradigm. J Biol Chem (2004) 279(2):1256-1261.
75. Omidfar K, Rasaee MJ, Modjtahedi H, Forouzandeh M, Taghikhani M, Golmakani N: Production of a novel camel single-domain antibody specifc for the type III mutant EGFR. Tumor Biol (2004) 25(5-6):296-305.
76. Saerens D, Kinne J, Bosmans E, Wernery U, Muyldermans S, Conrath K: Single domain antibodies derived from dromedary lymph node and peripheral blood lymphocytes sensing conformational variants of prostate-specifc antigen. J Biol Chem (2004) 279(50):51965-51972.
77. Conrath KE, Lauwereys M, Galleni M, Matagne A, Frere JM, Kinne J, Wyns L, Muyldermans S: β-Lactamase inhibitors derived from single-domain antibody fragments elicited in the Camelidae. Antimicrob Agents Chemother (2001) 45(10): 2807-2812.
78. Desmyter A, Spinelli S, Payan F, Lauwereys M, Wyns L, Muyldermans S, Cambillau C: Three camelid VHH domains in complex with porcine pancreatic α-amylase. Inhibition and versatility of binding topology. J Biol Chem (2002) 277(26): 23645-23650.
79. Transue TR, De Genst E, Ghahroudi MA, Wyns L, Muyldermans S: Camel single-domain antibody inhibits enzyme by mimicking carbohydrate substrate. Proteins (1998) 32(4):515-522.
80. De Genst E, Silence K, Decanniere K, Conrath K, Loris R, Kinne J, Muyldermans S, Wyns L: Molecular basis for the preferential cleft recognition by dromedary heavy-chain antibodies. Proc Natl Acad Sci U S A (2006) 103(12):4586-4591.
81. Sheriff S, Constantine KL: Redefning the minimal antigen-binding fragment. Nat Struct Biol (1996) 3(9):733-736.
82. Webster DM, Henry AH, Rees AR: Antibody-antigen interactions. Curr Opin Struct Biol (1994) 4(1):123-129.
83. Laskowski RA, Luscombe NM, Swindells MB, Thornton JM: Protein clefts in molecular recognition and function. Protein Sci (1996) 5(12):2438-2452.
84. Conrath EK, Lauwereys M, Wyns L, Muyldermans S: Camel single-domain antibodies as modular building units in bispecifc and bivalent antibody constructs. J Biol Chem (2001) 276(10):7346-7350.
85. Garaicoechea L, Olichon A, Marcoppido G, Wigdorovitz A, Mozgovoj M, Saif L, Surrey T, Parreno V: Llama-derived single-chain antibody fragments directed to rotavirus VP6 protein possess broad neutralizing activity in vitro and confer protection against diarrhea in mice. J Virol (2008) 82(19):9753-9764.
86. Groot AJ, Verheesen P, Westerlaken EJ, Gort EH, van der Groep P, Bovenschen N, van der Wall E, van Diest PJ, Shvarts A: Identifcation by phage display of single-domain antibody fragments specifc for the ODD domain in hypoxia-inducible factor 1α. Lab Invest (2006) 86(4):345-356.
87. Hmila I, Abdallah RB, Saerens D, Benlasfar Z, Conrath K, Ayeb ME, Muyldermans S, Bouhaouala-Zahar B: VHH, bivalent domains and chimeric heavy chain-only antibodies with high neutralizing effcacy for scorpion toxin AahI'. Mol Immunol (2008) 45(14): 3847-3856.
88. Mai KT, Perkins DG, Zhang J, Mackenzie CR: ES1, a new lung carcinoma antibody - An immunohistochemical study. Histopathology (2006) 49(5):515-522.
89. Stewart CS, MacKenzie CR, Hall JC: Isolation, characterization and pentamerization of ?-cobrotoxin specifc single-domain antibodies from a naïve phage display library: Preliminary fndings for antivenom development. Toxicon (2007) 49(5): 699-709.
90. Stone E, Hirama T, Chen W, Soltyk AL, Brunton J, MacKenzie CR, Zhang J: A novel pentamer versus pentamer approach to generating neutralizers of verotoxin 1. Mol Immunol (2007) 44(9):2487-2491.
91. Zhang J, Tanha J, Hirama T, Khieu NH, To R, Tong-Sevinc H, Stone E, Brisson JR, MacKenzie CR: Pentamerization of single-domain antibodies from phage libraries: A novel strategy for the rapid generation of high-avidity antibody reagents. J Mol Biol (2004) 335(1):49-56.
92. Stone E, Hirama T, Tanha J, Tong-Sevinc H, Li S, MacKenzie CR, Zhang J: The assembly of single domain antibodies into bispecifc decavalent molecules. J Immunol Methods (2007) 318(1-2):88-94.
93. Harmsen MM, Van Solt CB, Fijten HP, Van Setten MC: Prolonged in vivo residence times of llama single-domain antibody fragments in pigs by binding to porcine immunoglobulins. Vaccine (2005) 23(41):4926-4934.
94. Baral TN, Magez S, Stijlemans B, Conrath K, Vanhollebeke B, Pays E, Muyldermans S, De Baetselier P: Experimental therapy of African trypanosomiasis with a nanobody-conjugated human trypanolytic factor. Nat Med (2006) 12(5):580-584.
95. Szynol A, de Haard JJ, Veerman EC, de Soet JJ, van Nieuw Amerongen AV: Design of a peptibody consisting of the antimicrobial peptide Dhvar5 and a llama variable heavy-chain antibody fragment. Chem Biol Drug Des (2006) 67(6):425-431.
96. Kruger C, Hultberg A, Marcotte H, Hermans P, Bezemer S, Frenken LG, Hammarstrom L: Therapeutic effect of llama derived VHH fragments against Streptococcus mutans on the development of dental caries. Appl Microbiol Biotechnol (2006) 72(4):732-737.
97. Olichon A, Surrey T: Selection of genetically encoded fuorescent single domain antibodies engineered for effcient expression in Escherichia coli. J Biol Chem (2007) 282(50): 36314-36320.
98. van der Linden RH, Frenken LG, de Geus B, Harmsen MM, Ruuls RC, Stok W, de Ron L, Wilson S, Davis P, Verrips CT: Comparison of physical chemical properties of llama VHH antibody fragments and mouse monoclonal antibodies. Biochim Biophys Acta (1999) 1431(1):37-46.
99. Ewert S, Cambillau C, Conrath K, Pluckthun A: Biophysical properties of camelid VHH domains compared to those of human VH3 domains. Biochemistry (2002) 41(11):3628-3636.
100. Perez JM, Renisio JG, Prompers JJ, van Platerink CJ, Cambillau C, Darbon H, Frenken LG: Thermal unfolding of a llama antibody fragment: A two-state reversible process. Biochemistry (2001) 40(1):74-83.
101. Dolk E, van der Vaart M, Lutje Hulsik D, Vriend G, de Haard H, Spinelli S, Cambillau C, Frenken L, Verrips T: Isolation of llama antibody fragments for prevention of dandruff by phage display in shampoo. Appl Environ Microbiol (2005) 71(1):442-450.
102. Dumoulin M, Conrath K, Van Meirhaeghe A, Meersman F, Heremans K, Frenken LG, Muyldermans S, Wyns L, Matagne A: Single-domain antibody fragments with high conformational stability. Protein Sci (2002) 11(3):500-515. (Pubitemid 34171255)
103. Harmsen MM, van Solt CB, van Zijderveld-van Bemmel AM, Niewold TA, van Zijderveld FG: Selection and optimization of proteolytically stable llama single-domain antibody fragments for oral immunotherapy. Appl Microbiol Biotechnol (2006) 72(3):544-551.
104. Marcotte H, Pant N, Hammarström L: Engineered lactobody-producing lactobacilli: A novel form of therapy against rotavirus infection. Future Virol (2008) 3(4):327-341.
105. Pant N, Hultberg A, Zhao Y, Svensson L, Pan-Hammarstrom Q, Johansen K, Pouwels PH, Ruggeri FM, Hermans P, Frenken L, Boren T et al: Lactobacilli expressing variable domain of llama heavy-chain antibody fragments (lactobodies) confer protection against rotavirus-induced diarrhea. J Infect Dis (2006) 194(11):1580-1588.
106. van der Vaart JM, Pant N, Wolvers D, Bezemer S, Hermans PW, Bellamy K, Sarker SA, van der Logt CP, Svensson L, Verrips CT, Hammarstrom L et al: Reduction in morbidity of rotavirus induced diarrhoea in mice by yeast produced monovalent llama-derived antibody fragments. Vaccine (2006) 24(19): 4130-4137.
107. Ablynx NV: Ablynx announces technical advances in its nanobody platoform technology: Enhanced benefts from pulmonary delivery and half-life extension. Press Release (2009): January 14.
108. Deckers N, Saerens D, Kanobana K, Conrath K, Victor B, Wernery U, Vercruysse J, Muyldermans S, Dorny P: Nanobodies, a promising tool for species-specifc diagnosis of Taenia solium cysticercosis. Int J Parasitol (2008) 39(5):625-633.
109. Hulstein JJ, de Groot PG, Silence K, Veyradier A, Fijnheer R, Lenting PJ: A novel nanobody that detects the gain-of-function phenotype of von Willebrand factor in ADAMTS13 defciency and von Willebrand disease type 2B. Blood (2005) 106(9): 3035-3042.
110. Klooster R, Maassen BT, Stam JC, Hermans PW, Ten Haaft MR, Detmers FJ, de Haard HJ, Post JA, Theo Verrips C: Improved anti-IgG and HSA affnity ligands: Clinical application of VHH antibody technology. J Immunol Methods (2007) 324(1-2): 1-12.
111. Ladenson RC, Crimmins DL, Landt Y, Ladenson JH: Isolation and characterization of a thermally stable recombinant anti-caffeine heavy-chain antibody fragment. Anal Chem (2006) 78(13):4501-4508.
112. De Haard HJ, Bezemer S, Ledeboer AM, Muller WH, Boender PJ, Moineau S, Coppelmans MC, Verkleij AJ, Frenken LG, Verrips CT: Llama antibodies against a lactococcal protein located at the tip of the phage tail prevent phage infection. J Bacteriol (2005) 187(13):4531-4541.
113. Hultberg A, Tremblay DM, de Haard H, Verrips T, Moineau S, Hammarstrom L, Marcotte H: Lactobacillli expressing llama VHH fragments neutralise Lactococcus phages. BMC Biotechnol (2007) 7(58):1-7.
114. Ledeboer AM, Bezemer S, de Hiaard JJ, Schaffers IM, Verrips CT, van Vliet C, Dusterhoft EM, Zoon P, Moineau S, Frenken LG: Preventing phage lysis of Lactococcus lactis in cheese production using a neutralizing heavy-chain antibody fragment from llama. J Dairy Sci (2002) 85(6):1376-1382.
115. Harmsen MM, van Solt CB, Hoogendoorn A, van Zijderveld FG, Niewold TA, van der Meulen J: Escherichia coli F4 fmbriae specifc llama single-domain antibody fragments effectively inhibit bacterial adhesion in vitro but poorly protect against diarrhoea. Vet Microbiol (2005) 111(1-2):89-98.
116. Szynol A, de Soet JJ, Sieben-van Tuyl E, Bos JW, Frenken LG: Bactericidal effects of a fusion protein of llama heavy-chain antibodies coupled to glucose oxidase on oral bacteria. Antimicrob Agents Chemother (2004) 48(9):3390-3395.
117. Dekker S, Toussaint W, Panayotou G, de Wit T, Visser P, Grosveld F, Drabek D: Intracellularly expressed single-domain antibody against p15 matrix protein prevents the production of porcine retroviruses. J Virol (2003) 77(22):12132-12139.
118. Forsman A, Beirnaert E, Aasa-Chapman MM, Hoorelbeke B, Hijazi K, Koh W, Tack V, Szynol A, Kelly C, McKnight A, Verrips T et al: Llama antibody fragments with cross-subtype human immunodefciency virus type 1 (HIV-1)-neutralizing properties and high affnity for HIV-1 gp120. J Virol (2008) 82(24):12069-12081.
119. Serruys B, Van Houtte F, Verbrugghe P, Leroux-Roels G, Vanlandschoot P: Llama-derived single-domain intrabodies inhibit secretion of hepatitis B virions in mice. Hepatology (2008) 49(1):39-49.
120. Saerens D, Stijlemans B, Baral TN, Nguyen Thi GT, Wernery U, Magez S, De Baetselier P, Muyldermans S, Conrath K: Parallel selection of multiple anti-infectome nanobodies without access to purifed antigens. J Immunol Methods (2008) 329(1-2):138-150.
121. El Khattabi M, Adams H, Heezius E, Hermans P, Detmers F, Maassen B, van der Ley P, Tommassen J, Verrips T, Stam J: Llama single-chain antibody that blocks lipopolysaccharide binding and signaling: Prospects for therapeutic applications. Clin Vaccine Immunol (2006) 13(10):1079-1086.
122. Koch-Nolte F, Reyelt J, Schossow B, Schwarz N, Scheuplein F, Rothenburg S, Haag F, Alzogaray V, Cauerhff A, Goldbaum FA: Single domain antibodies from llama effectively and specifcally block T cell ecto-ADP-ribosyltransferase ART2.2 in vivo. Faseb J (2007) 21(13):3490-3498.
123. Meddeb-Mouelhi F, Bouhaouala-Zahar B, Benlasfar Z, Hammadi M, Mejri T, Moslah M, Karoui H, Khorchani T, El Ayeb M: Immunized camel sera and derived immunoglobulin subclasses neutralizing Androctonus australis hector scorpion toxins. Toxicon (2003) 42(7):785-791.
124. Kuan CT, Wikstrand CJ, Bigner DD: EGF mutant receptor vIII as a molecular target in cancer therapy. Endocr Relat Cancer (2001) 8(2):83-96.
125. Bakhtiari SH, Rahbarizadeh F, Hasannia S, Ahmadvand D, Iri-Sofa FJ, Rasaee MJ: Anti-MUC1 nanobody can redirect T-body cytotoxic effector function. Hybridoma (Larchmt) (2009) 28(2):85-92.
126. Rahbarizadeh F, Rasaee MJ, Forouzandeh M, Allameh A, Sarrami R, Nasiry H, Sadeghizadeh M: The production and characterization of novel heavy-chain antibodies against the tandem repeat region of MUC1 mucin. Immunol Invest (2005) 34(4):431-452.
127. Rahbarizadeh F, Rasaee MJ, Forouzandeh-Moghadam M, Allameh AA: High expression and purifcation of the recombinant camelid anti-MUC1 single domain antibodies in Escherichia coli. Protein Expr Purif (2005) 44(1):32-38.
128. McGonigal K, Tanha J, Palazov E, Li S, Gueorguieva-Owens D, Pandey S: Isolation and functional characterization of single domain antibody modulators of caspase-3 and apoptosis. Appl Biochem Biotechnol (2009) 157(2):226-236.
129. Groot AJ, Gort EH, van der Wall E, van Diest PJ, Vooijs M: Conditional inactivation of HIF-1 using intrabodies. Cell Oncol (2008) 30(5):397-409.
130. Burrows FJ, Derbyshire EJ, Tazzari PL, Amlot P, Gazdar AF, King SW, Letarte M, Vitetta ES, Thorpe PE: Up-regulation of endoglin on vascular endothelial cells in human solid tumors: Implications for diagnosis and therapy. Clin Cancer Res (1995) 1(12):1623-1634.
131. Ahmadvand D, Rasaee MJ, Rahbarizadeh F, Kontermann RE, Sheikholislami F: Cell selection and characterization of a novel human endothelial cell specifc nanobody. Mol Immunol (2009) 46(8-9):1814-1823.
132. Ahmadvand D, Rasaee MJ, Rahbarizadeh F, Mohammadi M: Production and characterization of a high-affnity nanobody against human endoglin. Hybridoma (Larchmt) (2008) 27(5): 353-360.
133. Behar G, Siberil S, Groulet A, Chames P, Pugniere M, Boix C, Sautes-Fridman C, Teillaud JL, Baty D: Isolation and characterization of anti-FcγRIII (CD16) llama single-domain antibodies that activate natural killer cells. Protein Eng Des Sel (2008) 21(1):1-10.
134. Muruganandam A, Tanha J, Narang S, Stanimirovic D: Selection of phage-displayed llama single-domain antibodies that transmigrate across human blood-brain barrier endothelium. Faseb J (2002) 16(2):240-242.
135. Gueorguieva D, Li S, Walsh N, Mukerji A, Tanha J, Pandey S: Identifcation of single-domain, Bax-specifc intrabodies that confer resistance to mammalian cells against oxidative-stress-induced apoptosis. Faseb J (2006) 20(14):2636-2638.
136. Habicht G, Haupt C, Friedrich RP, Hortschansky P, Sachse C, Meinhardt J, Wieligmann K, Gellermann GP, Brodhun M, Gotz J, Halbhuber KJ et al: Directed selection of a conformational antibody domain that prevents mature amyloid fbril formation by stabilizing Aβ protofbrils. Proc Natl Acad Sci USA (2007) 104(49):19232-19237.
137. Lafaye P, Achour I, England P, Duyckaerts C, Rougeon F: Single-domain antibodies recognize selectively small oligomeric forms of amyloid beta, prevent Aβ-induced neurotoxicity and inhibit fbril formation. Mol Immunol (2009) 46(4):695-704.
138. Dumoulin M, Last AM, Desmyter A, Decanniere K, Canet D, Larsson G, Spencer A, Archer DB, Sasse J, Muyldermans S, Wyns L et al: A camelid antibody fragment inhibits the formation of amyloid fbrils by human lysozyme. Nature (2003) 424(6950): 783-788.
139. van Koningsbruggen S, de Haard H, de Kievit P, Dirks RW, van Remoortere A, Groot AJ, van Engelen BG, den Dunnen JT, Verrips CT, Frants RR, van der Maarel SM: Llama-derived phage display antibodies in the dissection of the human disease oculopharyngeal muscular dystrophy. J Immunol Methods (2003) 279(1-2):149-161.
140. Chan PH, Pardon E, Menzer L, De Genst E, Kumita JR, Christodoulou J, Saerens D, Brans A, Bouillenne F, Archer DB, Robinson CV et al: Engineering a camelid antibody fragment that binds to the active site of human lysozyme and inhibits its conversion into amyloid fbrils. Biochemistry (2008) 47(42):11041-11054.
141. Spiel AO, Gilbert JC, Jilma B: von Willebrand factor in cardiovascular disease: Focus on acute coronary syndromes. Circulation (2008) 117(11):1449-1459.
142. Ulrichts H, Schoolmeester A, Hoefman S, Lauwereys M, Casteels P, Stanssens P, Baumeister J, Roodt J, de Jaegere PPT, Holz J: Anti-thrombotic drug candidate ALX-0081 shows superior preclinical effcacy and safety compared to current marketed antiplatelet drugs. J Thromb Haemost (2009) 7(s2): Abs AS-TH-024.
143. Holz J, Bartunek J, Barbato E, VerCruysse K, Pullan S, Heyndrickxs G: ALX-0081 a novel anti-thrombotic: First results of a multiple dose phase 1 study in patients with stable angina undergoing PCI. J Thromb Haemost (2009) 7(s2):Abs PP-WE-416.
144. Ablynx NV: Ablynx initiated phase II clinical trial for ALX-0081. Press Release (2009): September 01.
145. Ablynx NV: Ablynx announces positive phase I results for subcutaneous administration of its anti-thrombotic nanobody (ALX-0681). Press Release (2009): August 19.
146. Ablynx NV: Wyeth Pharmaceuticals initiates phase II and pays ablynx another milestone in the TNF nanobody collaboration. Press Release (2009): September 30.
147. Ablynx NV: Ablynx provides an R&D update announcing it has
148. Kabat EA, Wu TT, Perry HM, Gottesman KS, Foeller S: Sequences of Proteins of Immunological Interest. NIH, Bethesda, MD, USA (1991).
149. Abulrob A, Sprong H, Van Bergen en Henegouwen P, Stanimirovic D: The blood-brain barrier transmigrating single domain antibody: Mechanisms of transport and antigenic epitopes in human brain endothelial cells. J Neurochem (2005) 95(4):1201-1214.
150. Pipeline: Ablynx NV, Ghent/Zwijnaarde, Belgium (2008). www.ablynx.com/research/pipeline.htm
151. National Research Council of Canada (Muruganandam A, Tanha J, Narang S, Stanimirovic D): Single-domain brain-targeting antibody fragments derived from Llama antibodies. WO-02057445 (2002).