USOO922.1882B2 (12) United States Patent (10) Patent No.: US 9.221,882 B2 Skerra et al. (45) Date of Patent: Dec. 29, 2015 (54) BIOSYNTHETIC PROLINE/ALANINE WO WO 2007/103,515 A2 9, 2007 RANDOMCOL POLYPEPTDES AND THEIR WO WO 2008, 145400 A2 12/2008 USES WO WO 2010/091122 A1 8, 2010 (75) Inventors: Arne Skerra, Freising (DE); Uli Binder, OTHER PUBLICATIONS Freising (DE); Martin Schlapschy, Rudinger, Peptide Hormones, JA Parsons, Ed., 1976, pp. 1-7.* Freising (DE) SIGMA, 2004, pp. 1-2.* Berendsen, A Glimpae of the Holy Grail?, Science, 1998, 282, pp. (73) Assignees: Technische Universitat Munchen, 642-643. Munich (DE); XL-Protein GmbH, Ngo et al. Computational Complexity, Protein Structure Protection, Freising (DE) and the Levinthal Paradox, 1994, pp. 491-494.* (*) Notice: Subject to any disclaimer, the term of this Bradley et al., Limits of Cooperativity in a Structurally Modular patent is extended or adjusted under 35 Protein: Response of the Notch Ankyrin Domain to Analogous Alanine Substitutions in Each Repeat, J. Mol. BIoL (2002) 324. U.S.C. 154(b) by 0 days. 373-386. Voet et al. Biochemistry, John Wiley & Sons Inc., 1995, pp. 235 (21) Appl. No.: 13/697,569 241.* UniProt Protein Database, Very large tegument protein, UL36, pro (22) PCT Filed: May 20, 2011 tein accession Q7T5D9, accessed on Nov. 25, 2014.* International Search Report received in the parent application PCT/ (86). 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C07K I4/00 (2006.01) Breustedt, et al., The 1.8-A crystal structure of human tear lipocalin C07K 7/08 (2006.01) reveals an extended branched cavity with capacity for multiple (52) U.S. Cl. ligands. J Biol Chem, 2005, 280,484-93. CPC. C07K 14/00 (2013.01); C07K 7/08 (2013.01); Breustedt, et al...) Comparative ligand-binding analysis often human lipocalins. Biochim Biophys Acta, 2006, 1764, 161-73. C07K 14/001 (2013.01) Buscaglia, C.A., et al., “Tandem Amino Acid Repeats From Trypano (58) Field of Classification Search some cruzi Shed Antigens Increase the Half-Life of Proteins in CPC ........ C07K 7/08; C07K 14/00; C07K 14/001; Blood.” Blood, The American Society of Hematology, 1999, pp. C07K 17/00; C07K 147/08 2025-2032, vol. 93, No. 6. See application file for complete search history. Buscaglia, C.A., et al., “The Repetitive Domain of Trypanosoma Cruzi trans-Sialidase Enhances the Immune Response against the (56) References Cited Catalytic Domain.” The Journal of Infectious Diseases, 1998, pp. 431-436, vol. 177. U.S. PATENT DOCUMENTS (Continued) 5,677,172 A 10, 1997 Makarow Primary Examiner — James H Alstrum Acevedo 2003. O19074.0 A1 10, 2003 Altman Assistant Examiner — Erinne Dabkowski 2006/0252120 A1* 11/2006 Kieliszewski ............... 435/69.1 2009,0192087 A1 7/2009 Glass et al. (74) Attorney, Agent, or Firm — Foley & Lardner LLP 2010, O292130 A1 11/2010 Skerra (57) ABSTRACT We discovered that a biosynthetic polypeptide, consisting of FOREIGN PATENT DOCUMENTS at least about 50 proline (Pro) and alanine (Ala) amino acid residues, forms a random coil, and increases the stability of EP 1739.167 A1 1, 2007 other compounds to which it is conjugated. Such as Small EP 2173890 B1 3, 2011 JP 2007-529522 10/2007 molecules and polypeptides. 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