RRC ID 43462
Author Nogi T, Sangawa T, Tabata S, Nagae M, Tamura-Kawakami K, Beppu A, Hattori M, Yasui N, Takagi J.
Title Novel affinity tag system using structurally defined antibody-tag interaction: application to single-step protein purification.
Journal Protein Sci.
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.
Volume 17(12)
Pages 2120-6
Published 2008-12
DOI 10.1110/ps.038299.108
PII ps.038299.108
PMID 18787202
PMC PMC2590912
MeSH Animals Antibodies, Monoclonal / chemistry Antibodies, Monoclonal / immunology Antibodies, Monoclonal / metabolism* Antibody Affinity Cell Adhesion Molecules, Neuronal / genetics Cell Adhesion Molecules, Neuronal / isolation & purification Cell Line, Tumor Chromatography, Affinity / methods Crystallography, X-Ray / methods Epitopes / metabolism* Extracellular Matrix Proteins / genetics Extracellular Matrix Proteins / isolation & purification Humans Immunoglobulin Fab Fragments / chemistry Immunoglobulin Fab Fragments / genetics Immunoglobulin Fab Fragments / immunology Kinetics Mice Nerve Tissue Proteins / genetics Nerve Tissue Proteins / isolation & purification Peptides / chemistry Peptides / genetics Peptides / immunology Protein Binding Proteins / genetics Proteins / isolation & purification* Recombinant Fusion Proteins / genetics Recombinant Fusion Proteins / immunology Recombinant Fusion Proteins / isolation & purification Serine Endopeptidases / genetics Serine Endopeptidases / isolation & purification Structure-Activity Relationship
IF 2.41
Times Cited 14
Human and Animal Cells