title: Unnatural amino acid incorporation onto adenoviral (Ad) coat proteins facilitates chemoselective modification and retargeting of Ad type 5 vectors.
abstract: Surface modification of adenovirus vectors can improve tissue-selective targeting, attenuate immunogenicity, and enable imaging of particle biodistribution, thus significantly improving therapeutic potential. Currently, surface engineering is constrained by a combination of factors, including impact on viral fitness, limited access to functionality, or incomplete control over the site of modification. Here, we report a two-step labeling process involving an initial metabolic placement of a uniquely reactive unnatural amino acid, azidohomoalanine (Aha), followed by highly specific chemical modification. As genetic modification of adenovirus is unnecessary, vector production is exceedingly straightforward. Aha incorporation demonstrated no discernible impact on either virus production or infectivity of the resultant particles. "Click" chemical modification of surface-exposed azides was highly selective, allowing for the attachment of a wide range of functionality. Decoration of human adenovirus type 5 (hAd5) with folate, a known cancer-targeting moiety, provided an ∼20-fold increase in infection of murine breast cancer cells (4T1) in a folate receptor-dependent manner. This study demonstrates that incorporation of unnatural amino acids can provide a flexible, straightforward route for the selective chemical modification of adenoviral vectors.
authors: Department of Chemistry, State University of New York at Stony Brook, 1 John Toll Road, Stony Brook, NY 11794-3400, USA.
authors: Partha Sarathi
authors: PS
authors: Banerjee
authors: Philomena
authors: P
authors: Ostapchuk
authors: Patrick
authors: P
authors: Hearing
authors: Isaac Sheridan
authors: IS
authors: Carrico
references: Mol Ther. 2003 Oct;8(4):688-700
references: 14529842
references: Mol Ther. 2005 Jul;12(1):107-17
references: 15963926
references: Nat Biotechnol. 2000 Jul;18(7):723-7
references: 10888838
references: Curr Opin Chem Biol. 2009 Jun;13(3):256-62
references: 19419901
references: J Gen Virol. 2006 Nov;87(Pt 11):3151-3160
references: 17030847
references: Mol Ther. 2004 Jun;9(6):942-54
references: 15194061
references: Anal Biochem. 2001 Oct 1;297(1):1-9
references: 11567521
references: Bioconjug Chem. 2006 May-Jun;17(3):721-7
references: 16704210
references: J Inorg Biochem. 2004 Oct;98(10):1625-33
references: 15458825
references: Bioconjug Chem. 2005 Nov-Dec;16(6):1572-9
references: 16287257
references: J Virol. 2003 Sep;77(17):9553-66
references: 12915569
references: Gene Ther. 2007 Apr;14(8):627-37
references: 17251987
references: J Virol. 2006 Dec;80(24):12049-59
references: 17005667
references: J Virol. 2006 Dec;80(23):11881-6
references: 16987967
references: EMBO J. 2005 May 4;24(9):1645-54
references: 15861131
references: J Virol. 2009 Jan;83(2):1135-9
references: 19004948
references: Anal Biochem. 1999 Oct 15;274(2):283-8
references: 10527527
references: J Virol. 2004 Nov;78(22):12320-32
references: 15507619
references: Nat Rev Genet. 2007 Aug;8(8):573-87
references: 17607305
references: J Virol. 1997 Nov;71(11):8221-9
references: 9343173
references: J Virol. 2002 Sep;76(17):8621-31
references: 12163581
references: Virol J. 2008 Aug 21;5:98
references: 18718011
references: Cancer Gene Ther. 2006 Sep;13(9):830-44
references: 16439993
references: J Virol. 1998 Mar;72(3):1844-52
references: 9499035
references: Mol Ther. 2008 Jan;16(1):16-29
references: 17912234
references: Nat Biotechnol. 1996 Nov;14(11):1574-8
references: 9634824
references: Methods Mol Biol. 2009;542:57-74
references: 19565895
references: Methods Mol Biol. 2008;434:113-32
references: 18470642
references: Vaccine. 2006 Jan 30;24(5):671-82
references: 16154247
references: J Am Chem Soc. 2010 Oct 6;132(39):13615-7
references: 20831164
references: J Virol. 2003 Jul;77(13):7225-35
references: 12805421
references: J Mol Biol. 2005 Jun 10;349(3):526-37
references: 15890367
references: J Gene Med. 2002 Jul-Aug;4(4):356-70
references: 12124978
references: J Mol Biol. 2008 Nov 21;383(4):923-34
references: 18786542
references: Proc Natl Acad Sci U S A. 2002 Jan 8;99(1):19-24
references: 11752401
references: Virology. 2009 Feb 20;384(2):380-8
references: 19019405
references: J Control Release. 2007 Nov 6;123(2):164-71
references: 17854941