title: Tuning the serum persistence of human serum albumin domain III:diabody fusion proteins.
abstract: The long circulation persistence of human serum albumin (HSA) is enabled by its domain III (DIII) interaction with the neonatal Fc receptor (FcRn). A protein scaffold based on HSA DIII was designed. To modify the serum half life of the scaffold, residues H535, H510, and H464 were individually mutated to alanine. HSA DIII wild type (WT) and variants were fused to the anti-carcinoembryonic antigen (CEA) T84.66 diabody (Db), radiolabeled with (124)I and injected into xenografted athymic mice for serial PET/CT imaging. All proteins targeted the CEA-positive tumor. The mean residence times (MRT) of the proteins, calculated by quantifying blood activity from the PET images, were: Db-DIII WT (56.7 h), H535A (25 h), H510A (20 h), H464A (17 h), compared with Db (2.9 h). Biodistribution confirmed the order of blood clearance from slow to fast: Db-DIII WT > H535A > H510A > H464A > Db with 4.0, 2.0, 1.8, 1.6 and 0.08 %ID/g of remaining blood activity at 51 h, respectively. This study demonstrates that attenuating the DIII-FcRn interaction provides a way of controlling the pharmacokinetics of the entire Db-DIII fusion protein without compromising tumor targeting. H464 appears to be most crucial for FcRn binding (greatest reduction in MRT), followed by H510 and H535. By mutating the DIII scaffold, we can dial serum kinetics for imaging or therapy applications.
authors.affiliation: Crump Institute for Molecular Imaging, Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, California Nanosystems Institute, University of California, Los Angeles, 570 Westwood Plaza, CA 90095-1770, USA.
authors.forename: Vania E
authors.initials: VE
authors.lastname: Kenanova
authors.forename: Tove
authors.initials: T
authors.lastname: Olafsen
authors.forename: Felix B
authors.initials: FB
authors.lastname: Salazar
authors.forename: Lawrence E
authors.initials: LE
authors.lastname: Williams
authors.forename: Scott
authors.initials: S
authors.lastname: Knowles
authors.forename: Anna M
authors.initials: AM
authors.lastname: Wu
mesh_terms: D000409:Alanine / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D002272:Carcinoembryonic Antigen / Q000097:blood / Q000378:metabolism; D015395:Histocompatibility Antigens Class I / Q000097:blood / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007457:Iodine Radioisotopes; D007553:Isotope Labeling; D007700:Kinetics; D051379:Mice; D008819:Mice, Nude; D008969:Molecular Sequence Data; D009154:Mutation; D049268:Positron-Emission Tomography; D011961:Receptors, Fc / Q000097:blood / Q000737:chemistry / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000097:blood / Q000378:metabolism*; D012709:Serum Albumin / Q000737:chemistry / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D014057:Tomography, X-Ray Computed; D014183:Transplantation, Heterologous
chemical_list: D002272:Carcinoembryonic Antigen; D015395:Histocompatibility Antigens Class I; D007457:Iodine Radioisotopes; D011961:Receptors, Fc; D011993:Recombinant Fusion Proteins; D012709:Serum Albumin; D000409:Alanine; C100688:Fc receptor, neonatal
references.citation: Comput Programs Biomed. 1979 Mar;9(2):115-34
references.pmid: 761456
references.citation: Bioconjug Chem. 2001 Mar-Apr;12(2):220-8
references.pmid: 11312683
references.citation: Mol Cell. 2001 Apr;7(4):867-77
references.pmid: 11336709
references.citation: Cancer Immunol Immunother. 2005 Jun;54(6):535-47
references.pmid: 15592670
references.citation: Curr Opin Investig Drugs. 2009 Jun;10(6):572-8
references.pmid: 19513946
references.citation: J Immunol. 1997 Mar 1;158(5):2211-7
references.pmid: 9036967
references.citation: J Biol Chem. 2002 Nov 15;277(46):44050-60
references.pmid: 12213823
references.citation: J Biol Chem. 2001 Mar 2;276(9):6591-604
references.pmid: 11096108
references.citation: Nucl Med Biol. 2008 Feb;35(2):151-8
references.pmid: 18312824
references.citation: Proc Natl Acad Sci U S A. 1982 Jan;79(1):71-5
references.pmid: 6275391
references.citation: J Mol Biol. 2003 Feb 14;326(2):341-51
references.pmid: 12559905
references.citation: Drug Discov Today. 2009 Mar;14(5-6):278-83
references.pmid: 19073276
references.citation: Protein Eng. 1999 Jun;12(6):439-46
references.pmid: 10388840
references.citation: J Pharmacol Exp Ther. 2002 Nov;303(2):540-8
references.pmid: 12388634
references.citation: Traffic. 2009 May;10(5):600-14
references.pmid: 19192244
references.citation: Mol Imaging Biol. 2007 Sep-Oct;9(5):267-77
references.pmid: 17577599
references.citation: J Biol Chem. 2007 Apr 27;282(17):12650-60
references.pmid: 17347147
references.citation: Cancer Res. 1990 Apr 1;50(7):2128-34
references.pmid: 2107969
references.citation: Cancer Res. 2005 Jan 15;65(2):622-31
references.pmid: 15695407
references.citation: Biotechnology (N Y). 1992 Feb;10(2):169-75
references.pmid: 1369477
references.citation: Eur J Immunol. 1999 Sep;29(9):2819-25
references.pmid: 10508256
references.citation: Clin Cancer Res. 2003 May;9(5):1917-26
references.pmid: 12738750
references.citation: Proteins. 2003 Jul 1;52(1):80-7
references.pmid: 12784371
references.citation: Int J Cancer. 2001 Jun 1;92(5):718-24
references.pmid: 11340578
references.citation: Protein Sci. 1999 Jul;8(7):1423-31
references.pmid: 10422830
references.citation: Mol Imaging. 2003 Jul;2(3):131-7
references.pmid: 14649056
references.citation: Protein Eng Des Sel. 2006 Oct;19(10):453-60
references.pmid: 16882674
references.citation: J Exp Med. 2003 Feb 3;197(3):315-22
references.pmid: 12566415
references.citation: Biochemistry. 2006 Apr 18;45(15):4983-90
references.pmid: 16605266
references.citation: Mol Pharm. 2007 Sep-Oct;4(5):631-51
references.pmid: 17880166
references.citation: Nat Protoc. 2006;1(4):2048-60
references.pmid: 17487194
references.citation: J Med Chem. 2000 Apr 6;43(7):1253-6
references.pmid: 10753462
references.citation: J Biol Chem. 1989 Jan 15;264(2):953-9
references.pmid: 2910873
references.citation: J Biomol NMR. 1995 Sep;6(2):141-52
references.pmid: 8589603
references.citation: IEEE Trans Med Imaging. 1997 Apr;16(2):145-58
references.pmid: 9101324
references.citation: Eur J Immunol. 2006 Nov;36(11):3044-51
references.pmid: 17048273
references.citation: Nat Struct Biol. 1998 Sep;5(9):827-35
references.pmid: 9731778