title: Novel affinity tag system using structurally defined antibody-tag interaction: application to single-step protein purification.
abstract: Biologically important human proteins often require mammalian cell expression for structural studies, presenting technical and economical problems in the production/purification processes. We introduce a novel affinity peptide tagging system that uses a low affinity anti-peptide monoclonal antibody. Concatenation of the short recognition sequence enabled the successful engineering of an 18-residue affinity tag with ideal solution binding kinetics, providing a low-cost purification means when combined with nondenaturing elution by water-miscible organic solvents. Three-dimensional information provides a firm structural basis for the antibody-peptide interaction, opening opportunities for further improvements/modifications.
authors.affiliation: Laboratory of Protein Synthesis and Expression, Institute for Protein Research, Osaka University, Suita, Osaka 565-0871, Japan.
authors.forename: Terukazu
authors.initials: T
authors.lastname: Nogi
authors.forename: Takeshi
authors.initials: T
authors.lastname: Sangawa
authors.forename: Sanae
authors.initials: S
authors.lastname: Tabata
authors.forename: Masamichi
authors.initials: M
authors.lastname: Nagae
authors.forename: Keiko
authors.initials: K
authors.lastname: Tamura-Kawakami
authors.forename: Ayako
authors.initials: A
authors.lastname: Beppu
authors.forename: Mitsuharu
authors.initials: M
authors.lastname: Hattori
authors.forename: Norihisa
authors.initials: N
authors.lastname: Yasui
authors.forename: Junichi
authors.initials: J
authors.lastname: Takagi
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000276:immunology / Q000378:metabolism*; D000915:Antibody Affinity; D015816:Cell Adhesion Molecules, Neuronal / Q000235:genetics / Q000302:isolation & purification; D045744:Cell Line, Tumor; D002846:Chromatography, Affinity / Q000379:methods; D018360:Crystallography, X-Ray / Q000379:methods; D000939:Epitopes / Q000378:metabolism*; D016326:Extracellular Matrix Proteins / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D007700:Kinetics; D051379:Mice; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000302:isolation & purification; D010455:Peptides / Q000737:chemistry / Q000235:genetics / Q000276:immunology; D011485:Protein Binding; D011506:Proteins / Q000235:genetics / Q000302:isolation & purification*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D000091042:Reelin Protein; D012697:Serine Endopeptidases / Q000235:genetics / Q000302:isolation & purification; D013329:Structure-Activity Relationship
chemical_list: D000911:Antibodies, Monoclonal; D015816:Cell Adhesion Molecules, Neuronal; D000939:Epitopes; D016326:Extracellular Matrix Proteins; D007140:Immunoglobulin Fab Fragments; D009419:Nerve Tissue Proteins; D010455:Peptides; D011506:Proteins; D011993:Recombinant Fusion Proteins; D000091042:Reelin Protein; C429104:SPON1 protein, human; D012697:Serine Endopeptidases
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