Alternative protein scaffolds for molecular imaging and therapy

Engineering in Translational Medicine

Volumn , Issue , 2014, Pages 343-364

  Hackel, Benjamin J ^a  

^a University of Minnesota   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

CLINICAL RESEARCH; MOLECULAR IMAGING; SCAFFOLDS (BIOLOGY);

BIOLOGICAL RESPONSE; BIOPHYSICAL CHARACTERISTICS; FUSION PROTEINS; LOCAL DIVERSITIES; MOLECULAR FRAMEWORKS; MOLECULAR TARGETING; MOLECULAR TARGETS; PATHOPHYSIOLOGY; PROTEIN SCAFFOLDS; SPECIFIC BINDING;

ANTIBODIES;

EID: 84949179139     PISSN: None     EISSN: None     Source Type: Book    
DOI: 10.1007/978-1-4471-4372-7_13     Document Type: Chapter

References (103)

1. Scott AM, Wolchok JD, Old LJ (2012) Antibody therapy of cancer. Nat Rev Cancer 12: 278-287. doi: 10.1038/nrc3236
2. Yuan F, Dellian M, Fukumura D et al (1995) Vascular permeability in a human tumor xenograft: molecular size dependence and cutoff size. Cancer Res 55: 3752-3756
3. Schmidt MM, Wittrup KD (2009) A modeling analysis of the effects of molecular size and binding affinity on tumor targeting. Mol Cancer Ther 8: 2861-2871. doi: 10.1158/1535-7163.MCT-09-0195
4. Binz HK, Amstutz P, Plückthun A (2005) Engineering novel binding proteins from nonimmunoglobulin domains. Nat Biotechnol 23: 1257-1268. doi: 10.1038/nbt1127
5. Löfblom J, Frejd FY, Ståhl S (2011) Non-immunoglobulin based protein scaffolds. Curr Opin Biotechnol. doi: 10.1016/j.copbio.2011.06.002
6. Löfblom J, Feldwisch J, Tolmachev V et al (2010) Affibody molecules: engineered proteins for therapeutic, diagnostic and biotechnological applications. FEBS Lett 584: 2670-2680. doi: 10.1016/j.febslet.2010.04.014
7. Lipovsek D (2011) Adnectins: engineered target-binding protein therapeutics. Protein Eng Des Sel 24: 3-9. doi: 10.1093/protein/gzq097
8. Tamaskovic R, Simon M, Stefan N et al (2012) Designed ankyrin repeat proteins (DARPins) from research to therapy. Meth Enzymol 503: 101-134. doi: 10.1016/B978-0-12-396962-0.00005-7
9. Gebauer M, Skerra A (2012) Anticalins small engineered binding proteins based on the lipocalin scaffold. Meth Enzymol 503: 157-188. doi: 10.1016/B978-0-12-396962-0.00007-0
10. Moore SJ, Leung CL, Cochran Jr. (2012) Knottins: disulfide-bonded therapeutic and diagnostic peptides. Drug Discov Today: Technol 9: e3-e11. doi: 10.1016/j.ddtec. 2011.07.003
11. Ståhl S, Kronqvist N, Jonsson A, Löfblom J (2012) Affinity proteins and their generation. J Chem Technol Biotechnol 88: 25-38. doi: 10.1002/jctb.3929
12. Gebauer M, Skerra A (2009) Engineered protein scaffolds as next-generation antibody therapeutics. Curr Opin Chem Biol 13: 245-255. doi: 10.1016/j.cbpa.2009.04.627
13. Hackel BJ, Kapila A, Wittrup KD (2008) Picomolar affinity fibronectin domains engineered utilizing loop length diversity, recursive mutagenesis, and loop shuffling. J Mol Biol 381: 1238-1252. doi: 10.1016/j.jmb.2008.06.051
14. Batori V, Koide A, Koide S (2002) Exploring the potential of the monobody scaffold: effects of loop elongation on the stability of a fibronectin type III domain. Protein Eng 15: 1015-1020
15. Parker MH, Chen Y, Danehy F et al (2005) Antibody mimics based on human fibronectin type three domain engineered for thermostability and high-Affinity binding to vascular endothelial growth factor receptor two. Protein Eng Des Sel 18: 435-444. doi: 10.1093/protein/gzi050
16. Hackel BJ, Wittrup K.D. 2010 The full amino acid repertoire is superior to serine/tyrosine for selection of high affinity immunoglobulin G binders from the fibronectin scaffold. Protein Eng Des Sel 23: 211-219. doi: 10.1093/protein/gzp083
17. Hackel BJ Ackerman ME Howland SW Wittrup KD. 2010 Stability and CDR composition biases enrich binder functionality landscapes J Mol Biol 401 84-96 doi: 10.1016/j.jmb.2010.06.004
18. Koide A, Jordan MR, Horner SR et al (2001) Stabilization of a fibronectin type III domain by the removal of unfavorable electrostatic interactions on the protein surface. Biochemistry 40: 10326-10333
19. Getmanova EV Chen Y Bloom L. et al. 2006 Antagonists to human and mouse vascular endothelial growth factor receptor 2 generated by directed protein evolution in vitro Chem Biol 13 549-556 doi: 10.1016/j.chembiol.2005.12.009
20. Mamluk R, Carvajal IM, Morse BA et al (2010) Anti-tumor effect of CT-322 as an adnectin inhibitor of vascular endothelial growth factor receptor-2. mAbs, vol 2, pp 199-208
21. Dineen SP, Sullivan LA, Beck AW et al (2008) The Adnectin CT-322 is a novel VEGF receptor 2 inhibitor that decreases tumor burden in an orthotopic mouse model of pancreatic cancer. BMC Cancer 8: 352. doi: 10.1186/1471-2407-8-352
22. Ackermann M, Carvajal IM, Morse BA et al (2011) Adnectin CT-322 inhibits tumor growth and affects microvascular architecture and function in Colo205 tumor xenografts. Int J Oncol 38: 71-80. doi: 10.3892/ijo-00000825
23. Waters JD, Sanchez C, Sahin A et al (2012) CT322, a VEGFR-2 antagonist, demonstrates anti-glioma efficacy in orthotopic brain tumor model as a single agent or in combination with temozolomide and radiation therapy. J Neurooncol 110: 37-48. doi: 10.1007/s11060-012-0948-7
24. Tolcher AW, Sweeney CJ, Papadopoulos K et al (2011) Phase I and pharmacokinetic study of CT-322 (BMS-844203), a targeted Adnectin inhibitor of VEGFR-2 based on a domain of human fibronectin. Clin Cancer Res 17: 363-371. doi: 10.1158/1078-0432.CCR-10-1411
25. Paschold EH, Mazieres J, Lena H et al (2012) A randomized, double-blinded, phase II study of paclitaxel/carboplatin (PC) plus CT-322 versus PC plus bevacizumab (Bev) as first-line treatment for advanced nonsquamous non-small cell lung cancer (NSCLC). J Clin Oncol 30: a7584
26. Ackermann M, Morse BA, Delventhal V et al (2012) Anti-VEGFR2 and anti-IGF-1RAdnectins inhibit Ewing's sarcoma A673-xenograft growth and normalize tumor vascular architecture. Angiogenesis 15: 685-695. doi: 10.1007/s10456-012-9294-9
27. Emanuel SL, Engle LJ, Chao G et al. (2011) A fibronectin scaffold approach to bispecific inhibitors of epidermal growth factor receptor and insulin-like growth factor-I receptor. mAbs vol 3, pp 38-48
28. Wang H, Wang L, Cao K et al (2012) Development of a carbon-14 labeling approach to support disposition studies with a pegylated biologic. Drug Metab Dispos 40: 1677-1685. doi: 10.1124/dmd.112.044792
29. Spangler JB, Manzari MT, Rosalia EK et al (2012) Triepitopic antibody fusions inhibit cetuximab-resistant BRAF and KRAS mutant tumors via EGFR signal repression. J Mol Biol 422: 532-544. doi: 10.1016/j.jmb.2012.06.014
30. Hackel BJ, Kimura RH, Gambhir SS (2012) Use of 64Cu-labeled fibronectin domain with EGFR-overexpressing tumor xenograft: molecular imaging. Radiology 263: 179-188. doi: 10.1148/radiol.12111504
31. Hackel BJ, Sathirachinda A, Gambhir SS (2012) Designed hydrophilic and charge mutations of the fibronectin domain: towards tailored protein biodistribution. Protein Eng Des Sel 25: 639-647. doi: 10.1093/protein/gzs036
32. Pirie CM, Hackel BJ, Rosenblum MG, Wittrup KD (2011) Convergent potency of internalized gelonin immunotoxins across varied cell lines, antigens, and targeting moieties. J Biol Chem 286: 4165-4172. doi: 10.1074/jbc.M110.186973
33. Brautbar A, Ballantyne CM (2011) Pharmacological strategies for lowering LDL cholesterol: statins and beyond. Nat Rev Cardiol 8: 253-265. doi: 10.1038/nrcardio.2011.2
34. Colgrave ML, Craik DJ (2004) Thermal, chemical, and enzymatic stability of the cyclotide kalata B1: the importance of the cyclic cystine knot. Biochemistry 43: 5965-5975. doi: 10.1021/bi049711q
35. Werle M, Schmitz T, Huang H-L et al (2006) The potential of cystine-knot microproteins as novel pharmacophoric scaffolds in oral peptide drug delivery. J Drug Target 14: 137-146. doi: 10.1080/10611860600648254
36. Moore SJ, Cochran Jr. (2012) Engineering knottins as novel binding agents. Meth Enzymol 503: 223-251. doi: 10.1016/B978-0-12-396962-0.00009-4
37. Gracy J, Le-Nguyen D, Gelly J-C et al (2008) KNOTTIN: the knottin or inhibitor cystine knot scaffold in 2007. Nucleic Acids Res 36: D314-D319. doi: 10.1093/nar/gkm939
38. Perez-Pinzon MA, Yenari MA, Sun GH et al (1997) SNX-111, a novel, presynaptic N-type calcium channel antagonist, is neuroprotective against focal cerebral ischemia in rabbits. J Neurol Sci 153: 25-31
39. Joppa MA, Gogas KR, Foster AC, Markison S (2007) Central infusion of the melanocortin receptor antagonist agouti-related peptide (AgRP(83-132)) prevents cachexia-related symptoms induced by radiation and colon-26 tumors in mice. Peptides 28: 636-642. doi: 10.1016/j.peptides.2006.11.021
40. Hockaday DC, Shen S, Fiveash J et al (2005) Imaging glioma extent with 131I-TM-601. J Nucl Med 46: 580-586
41. Mamelak AN (2006) Phase I single-dose study of intracavitary-Administered iodine-131-TM-601 in adults with recurrent high-grade glioma. J Clin Oncol 24: 3644-3650. doi: 10.1200/JCO.2005.05.4569
42. Mrugala MM, Adair JE, Kiem H-P (2012) Outside the box-novel therapeutic strategies for glioblastoma. Cancer J 18: 51-58. doi: 10.1097/PPO.0b013e318243f785
43. Veiseh M, Gabikian P, Bahrami S-B et al (2007) Tumor paint: a chlorotoxin: Cy5.5 bioconjugate for intraoperative visualization of cancer foci. Cancer Res 67: 6882-6888. doi: 10.1158/0008-5472.CAN-06-3948
44. Sun C Veiseh O Gunn J. et al. 2008 In vivo MRI detection of gliomas by chlorotoxinconjugated superparamagnetic nanoprobes. Small 4 372-379 doi: 10.1002/smll.200700784
45. Krause S, Schmoldt H-U, Wentzel A et al (2007) Grafting of thrombopoietin-mimetic peptides into cystine knot miniproteins yields high-Affinity thrombopoietin antagonists and agonists. FEBS J 274: 86-95. doi: 10.1111/j.1742-4658.2006.05567.x
46. Kimura RH, Levin AM, Cochran FV, Cochran Jr. (2009) Engineered cystine knot peptides that bind alphavbeta3, alphavbeta5, and alpha5beta1 integrins with low-nanomolar affinity. Proteins 77: 359-369. doi: 10.1002/prot.22441
47. Kimura RH, Cheng Z, Gambhir SS, Cochran Jr. (2009) Engineered knottin peptides: a new class of agents for imaging integrin expression in living subjects. Cancer Res 69: 2435-2442. doi: 10.1158/0008-5472.CAN-08-2495
48. Kimura RH, Miao Z, Cheng Z et al (2010) A dual-labeled knottin peptide for PET and nearinfrared fluorescence imaging of integrin expression in living subjects. Bioconjug Chem 21: 436-444. doi: 10.1021/bc9003102
49. Miao Z, Ren G, HHongguang L (2009) An engineered knottin peptide labeled with 18F for PET imaging of integrin expression. Bioconjug Chem 20: 2342-2347. doi: 10.1021/bc900361g
50. Nielsen CH, Kimura RH, Withofs N et al (2010) PET imaging of tumor neovascularization in a transgenic mouse model with a novel 64Cu-DOTA-knottin peptide. Cancer Res 70: 9022-9030. doi: 10.1158/0008-5472.CAN-10-1338
51. Willmann JK, Kimura RH, Deshpande N et al (2010) Targeted contrast-enhanced ultrasound imaging of tumor angiogenesis with contrast microbubbles conjugated to integrin-binding knottin peptides. J Nucl Med 51: 433-440. doi: 10.2967/jnumed.109.068007
52. Silverman AP, Levin AM, Lahti JL, Cochran Jr. (2009) Engineered cystine-knot peptides that bind alpha(v)beta(3) integrin with antibody-like affinities. J Mol Biol 385: 1064-1075. doi: 10.1016/j.jmb.2008.11.004
53. Jiang L, Kimura RH, Miao Z et al (2010) Evaluation of a (64)Cu-labeled cystine-knot peptide based on agouti-related protein for PET of tumors expressing alphavbeta3 integrin. J Nucl Med 51: 251-258. doi: 10.2967/jnumed.109.069831
54. Jiang L, Miao Z, Kimura RH et al (2012) 111In-labeled cystine-knot peptides based on the agouti-related protein for targeting tumor angiogenesis. J Biomed Biotechnol 2012: 1-8. doi: 10.1002/ijc.23575
55. Jiang H, Moore SJ, Liu S et al (2013) A novel radiofluorinated agouti-related protein for tumor angiogenesis imaging. Amino Acids 44: 673-681. doi: 10.1007/s00726-012-1391-y
56. Kimura RH, Teed R, Hackel BJ et al (2012) Pharmacokinetically stabilized cystine knot peptides that bind alpha-v-beta-6 integrin with single-digit nanomolar affinities for detection of pancreatic cancer. Clin Cancer Res 18: 839-849. doi: 10.1158/1078-0432.CCR-11-1116
57. Hackel BJ, Kimura RH, Miao Z et al (2013) 18F-labeled cystine knot peptides for PET imaging of integrin avb6. J Nucl Med 54(7): 1101-1105
58. Jiang L, Miao Z, Kimura RH et al (2011) Preliminary evaluation of (177)Lu-labeled knottin peptides for integrin receptor-targeted radionuclide therapy. Eur J Nucl Med Mol Imaging 38: 613-622. doi: 10.1007/s00259-010-1684-x
59. Kimura RH, Jones DS, Jiang L et al (2011) Functional mutation of multiple solvent-exposed loops in the Ecballium elaterium trypsin inhibitor-II cystine knot miniprotein. PLoS ONE 6: e16112. doi: 10.1371/journal.pone.0016112
60. Souriau C, Chiche L, Irving R, Hudson P (2005) New binding specificities derived from Min-23, a small cystine-stabilized peptidic scaffold. Biochemistry 44: 7143-7155. doi: 10.1021/bi0481592
61. Zahnd C, Wyler E, Schwenk JM et al (2007) A designed ankyrin repeat protein evolved to picomolar affinity to Her2. J Mol Biol 369: 1015-1028. doi: 10.1016/j.jmb.2007.03.028
62. Zahnd C, Kawe M, Stumpp MT et al (2010) Efficient tumor targeting with high-Affinity designed ankyrin repeat proteins: effects of affinity and molecular size. Cancer Res 70: 1595-1605. doi: 10.1158/0008-5472.CAN-09-2724
63. Theurillat J-P, Dreier B, Nagy-Davidescu G et al (2010) Designed ankyrin repeat proteins: a novel tool for testing epidermal growth factor receptor 2 expression in breast cancer. Mod Pathol 23: 1289-1297. doi: 10.1038/modpathol.2010.103
64. Münch RC, Mühlebach MD, Schaser T et al (2011) DARPins: an efficient targeting domain for lentiviral vectors. Mol Ther 19: 686-693. doi: 10.1038/mt.2010.298
65. Friedrich K, Hanauer JR, Prüfer S et al (2013) DARPin-targeting of measles virus: unique bispecificity, effective oncolysis, and enhanced safety. Mol Ther. doi: 10.1038/mt.2013.16
66. Martin-Killias P, Patricia M-K, Stefan N et al (2011) A novel fusion toxin derived from an EpCAM-specific designed ankyrin repeat protein has potent antitumor activity. Clin Cancer Res 17: 100-110. doi: 10.1158/1078-0432.CCR-10-1303
67. Wolf S, Souied EH, Mauget-Faysse M et al (2011) Phase I MP0112 wet AMD study: results of a single escalating dose study with DARPin MP0112 in wet AMD. 2011 annual meeting of the association for research in vision and ophthalmology, vol 1655
68. Campochiaro PA, Channa R, Berger BB et al (2013) Treatment of diabetic macular edema with a designed ankyrin repeat protein that binds vascular endothelial growth factor: a phase I/II study. Am J Ophthalmol 155(697-704): e2. doi: 10.1016/j.ajo.2012.09.032
69. Kim HJ, Eichinger A, Skerra A (2009) High-Affinity recognition of lanthanide (III) chelate complexes by a reprogrammed human lipocalin 2. J Am Chem Soc 131: 3565-3576. doi: 10.1021/ja806857r
70. Schönfeld D, Matschiner G, Chatwell L et al (2009) An engineered lipocalin specific for CTLA-4 reveals a combining site with structural and conformational features similar to antibodies. Proc Natl Acad Sci 106: 8198-8203. doi: 10.1073/pnas.0813399106
71. Nunn MA, Sharma A, Paesen GC et al (2005) Complement inhibitor of C5 activation from the soft tick Ornithodoros moubata. J Immunol 174: 2084-2091
72. Halstead SK, Humphreys PD, Zitman FMP et al (2008) C5 inhibitor rEV576 protects against neural injury in an in vitro mouse model of Miller Fisher syndrome. J Peripher Nerv Syst 13: 228-235. doi: 10.1111/j.1529-8027.2008.00181.x
73. Soltys J, Kusner LL, Young A et al (2009) Novel complement inhibitor limits severity of experimentally myasthenia gravis. Ann Neurol 65: 67-75. doi: 10.1002/ana.21536
74. Carrera-Marin AL, Romay-Penabad Z, Machin S et al (2011) C5 inhibitor rEV576 ameliorates in vivo effects of antiphospholipid antibodies. Arthritis and Rheumatism 63: S5-S5
75. Couillin I, Maillet I, Vargaftig BB et al (2004) Arthropod-derived histamine-binding protein prevents murine allergic asthma. J Immunol 173: 3281-3286
76. Ryffel B, Couillin I, Maillet I et al (2005) Histamine scavenging attenuates endotoxininduced acute lung injury. Ann N Y Acad Sci 1056: 197-205. doi: 10.1196/annals.1352.034
77. Hauptman PJ, Kelly RA (1999) Digitalis. Circulation 99: 1265-1270
78. Schlehuber S, Skerra A (2005) Lipocalins in drug discovery: from natural ligand-binding proteins to "anticalins". Drug Discov Today 10: 23-33. doi: 10.1016/S1359-6446(04)03294-5
79. Eyer F, Steimer W, Nitzsche T et al (2012) Intravenous application of an anticalin dramatically lowers plasma digoxin levels and reduces its toxic effects in rats. Toxicol Appl Pharmacol 263: 352-359. doi: 10.1016/j.taap.2012.07.009
80. Wurch T, Pierré A, Depil S (2012) Novel protein scaffolds as emerging therapeutic proteins: from discovery to clinical proof-of-concept. Trends Biotechnol 30: 575-582. doi: 10.1016/j.tibtech.2012.07.006
81. Mross K. Fischer R. Richly H. et al. 2011 Abstract A212: first in human phase I study of PRS-050 (Angiocal), a VEGF-A targeting anticalin, in patients with advanced solid tumors: results of a dose escalation study Mol Cancer Ther 10 A212-A212 doi: 10.1158/1535-7163.TARG-11-A212
82. Steiner M, Gutbrodt K, Krall N, Neri D (2013) Tumor-targeting antibody-Anticalin fusion proteins for in vivo pretargeting applications Bioconjug Chem 24: 234-241. doi: 10.1021/bc300567a
83. Lendel C, Dincbas-Renqvist V, Flores A et al (2004) Biophysical characterization of Z(SPA-1)-A phage-display selected binder to protein A. Protein Sci 13: 2078-2088. doi: 10.1110/ps.04728604
84. Feldwisch J, Tolmachev V, Lendel C et al (2010) Design of an optimized scaffold for affibody molecules. J Mol Biol 398: 232-247. doi: 10.1016/j.jmb.2010.03.002
85. Lundberg E, Brismar H, Gräslund T (2009) Selection and characterization of Affibody ligands to the transcription factor c-Jun. Biotechnol Appl Biochem 52: 17-27. doi: 10.1042/BA20070178
86. Nygren P-Å (2008) Alternative binding proteins: affibody binding proteins developed from a small three-helix bundle scaffold. FEBS J 275: 2668-2676. doi 10.1111/j.1742-4658.2008.06438.x
87. Lindborg M, Cortez E, Höidén-Guthenberg I et al (2011) Engineered high-Affinity affibody molecules targeting platelet-derived growth factor receptor b in vivo. J Mol Biol 407: 298-315. doi: 10.1016/j.jmb.2011.01.033
88. Kronqvist N, Malm M, Göstring L et al (2011) Combining phage and staphylococcal surface display for generation of ErbB3-specific Affibody molecules
89. Webster JM, Zhang R, Gambhir SS et al (2009) Engineered two-helix small proteins for molecular recognition ChemBioChem 10: 1293-1296. doi: 10.1002/cbic.200900062
90. Ren G, Zhang R, Liu Z et al (2009) A 2-helix small protein labeled with 68 Ga for PET imaging of HER2 expression. J Nucl Med 50: 1492-1499. doi: 10.2967/jnumed.109.064287
91. Miao Z, Ren G, Jiang L et al (2011) A novel (18)F-labeled two-helix scaffold protein for PET imaging of HER2-positive tumor. Eur J Nucl Med Mol Imaging. doi: 10.1007/s00259-011-1879-9
92. Ahlgren S, Tolmachev V (2010) Radionuclide molecular imaging using Affibody molecules. Curr Pharm Biotechnol 11: 581-589
93. Miao Z, Levi J, Cheng Z (2010) Protein scaffold-based molecular probes for cancer molecular imaging. Amino Acids. doi: 10.1007/s00726-010-0503-9
94. Yang M, Cheng K, Qi S et al (2013) Affibody modified and radiolabeled gold-Iron oxide hetero-nanostructures for tumor PET, optical and MR imaging. Biomaterials 34: 2796-2806. doi: 10.1016/j.biomaterials.2013.01.014
95. Baum RP, Prasad V, Müller D et al (2010) Molecular imaging of HER2-expressing malignant tumors in breast cancer patients using synthetic 111In- or 68 Ga-labeled affibody molecules. J Nucl Med 51: 892-897. doi: 10.2967/jnumed.109.073239
96. Zielinski R Lyakhov I Hassan M. et al. 2011 HER2-Affitoxin: a potent therapeutic agent for the treatment of HER2-overexpressing tumors Clin Cancer Res 17 5071-5081 doi: 10.1158/1078-0432.CCR-10-2887
97. Andersen JT, Pehrson R, Tolmachev V et al (2011) Extending half-life by indirect targeting of the neonatal Fc receptor (FcRn) using a minimal albumin binding domain. J Biol Chem 286: 5234-5241. doi: 10.1074/jbc.M110.164848
98. Steffen A-C, Almqvist Y, Chyan M-K et al (2007) Biodistribution of 211At labeled HER-2 binding affibody molecules in mice. Oncol Rep 17: 1141-1147
99. Myhre S, Henning P, Friedman M et al (2009) Re-targeted adenovirus vectors with dual specificity; binding specificities conferred by two different Affibody molecules in the fiber. Gene Ther 16: 252-261. doi: 10.1038/gt.2008.160
100. Lehmann A (2008) Ecallantide (DX-88), a plasma kallikrein inhibitor for the treatment of hereditary angioedema and the prevention of blood loss in on-pump cardiothoracic surgery. Expert Opin Biol Ther 8: 1187-1199. doi: 10.1517/14712598.8.8.1187
101. Zuraw B, Yasothan U, Kirkpatrick P (2010) Ecallantide. Nat Rev Drug Discov 9: 189-190. doi: 10.1038/nrd3125
102. Silverman J, Liu Q, Lu Q et al (2005) Multivalent avimer proteins evolved by exon shuffling of a family of human receptor domains. Nat Biotechnol 23: 1556-1561. doi: 10.1038/nbt1166
103. Hudson PJ, Souriau C (2003) Engineered antibodies. Nat Med 9: 129-134. doi: 10.1038/nm0103-129