title: Flexible antibodies with nonprotein hinges.
abstract: There is a significant need for antibodies that can bind targets with greater affinity. Here we describe a novel strategy employing chemical semisynthesis to produce symmetroadhesins: antibody-like molecules having nonprotein hinge regions that are more flexible and extendible and are capable of two-handed binding. Native chemical ligation was carried out under mild, non-denaturing conditions to join a ligand binding domain (Aβ peptide) to an IgG1 Fc dimer via discrete oxyethylene oligomers of various lengths. Two-handed Aβ-Fc fusion proteins were obtained in quantitative yield and shown by surface plasmon resonance to bind an anti-Aβ antibody with a K(D) at least two orders of magnitude greater than the cognate Aβ peptide. MALDI-TOF MS analysis confirmed the protein/nonprotein/protein structure of the two-handed molecules, demonstrating its power to characterize complex protein-nonprotein hybrids by virtue of desorption/ionization mediated by peptide sequences contained therein. We anticipate many applications for symmetroadhesins that combine the target specificity of antibodies with the novel physical, chemical and biological properties of nonprotein hinges.
authors.affiliation: Blood Systems Research Institute, San Francisco, CA 94118, USA. dcapon@alum.mit.edu
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Capon
authors.forename: Naoki
authors.initials: N
authors.lastname: Kaneko
authors.forename: Takayuki
authors.initials: T
authors.lastname: Yoshimori
authors.forename: Takashi
authors.initials: T
authors.lastname: Shimada
authors.forename: Florian M
authors.initials: FM
authors.lastname: Wurm
authors.forename: Peter K
authors.initials: PK
authors.lastname: Hwang
authors.forename: Xiaohe
authors.initials: X
authors.lastname: Tong
authors.forename: Staci A
authors.initials: SA
authors.lastname: Adams
authors.forename: Graham
authors.initials: G
authors.lastname: Simmons
authors.forename: Taka-Aki
authors.initials: TA
authors.lastname: Sato
authors.forename: Koichi
authors.initials: K
authors.lastname: Tanaka
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007700:Kinetics; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D053758:Nanoparticles / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry; D018583:Pliability; D011485:Protein Binding; D011487:Protein Conformation; D021122:Protein Subunits / Q000737:chemistry / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000138:chemical synthesis / Q000737:chemistry; D020349:Surface Plasmon Resonance; D014357:Trypsin / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D010455:Peptides; D021122:Protein Subunits; D011993:Recombinant Fusion Proteins; D014357:Trypsin
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