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Title: Bifunctional polyacrylamide based polymers for the specific binding of hexahistidine tagged proteins on gold surfaces

Journal Article · · Physical Chemistry Chemical Physics. PCCP
DOI: https://doi.org/10.1039/b920713a · OSTI ID:1875079
 [1];  [2];  [2]
  1. Univ. of Illinois at Urbana-Champaign, IL (United States); University of Illinois
  2. Univ. of Illinois at Urbana-Champaign, IL (United States)

We describe a modified bifunctional analogue of polyacrylamide that spontaneously forms self-assembled polymeric thin films on Au surfaces. The film is engineered to specifically bind histidine tagged proteins (6His), while simultaneously remaining inherently resistant to the non-specific adsorption of proteins in solution. The backbone of a polyacrylamide-co-n-acryloxysuccinimide copolymer is functionalized via tandem active ester (NHS) couplings with 3-(methylthio)propylamine (MTP) and nitrilotriacetic acid (NTA). The resulting functionalized polymers form stable and exceptionally hydrophilic thin films that are ~2–5 nm thick, a mass coverage that varies with the MTP graft density. These films are characterized using a variety of techniques (X-ray photoelectron spectroscopy (XPS), reflection absorption infrared spectroscopy (RAIRS), ellipsometry, surface plasmon resonance (SPR), and matrix assisted laser desorption ionization (MALDI)) to establish their structure and function. The protein resistance of the films, as demonstrated by their exposure to solutions of bovine serum albumin (BSA), can be modulated by the amount of MTP grafted to the polymer, which in turn, affects their mass coverage. We show that it is possible to specifically capture hexahistidine tagged proteins with low incidences of nonspecific adsorption using these materials, a discrimination quantified using surface plasmon resonance (SPR) at concentrations down to ~20 nM. Furthermore, these polymers also bind strongly to the surfaces of Au nanoparticles, stabilizing them against aggregation, providing them with a similar capacity to selectively bind 6His tagged proteins that can then be speciated using MALDI.

Research Organization:
Univ. of Illinois at Urbana-Champaign, IL (United States)
Sponsoring Organization:
National Science Foundation; USDOE Office of Science (SC), Basic Energy Sciences (BES). Materials Sciences & Engineering Division
Grant/Contract Number:
FG02-07ER46471
OSTI ID:
1875079
Journal Information:
Physical Chemistry Chemical Physics. PCCP, Journal Name: Physical Chemistry Chemical Physics. PCCP Journal Issue: 17 Vol. 12; ISSN 1463-9076
Publisher:
Royal Society of ChemistryCopyright Statement
Country of Publication:
United States
Language:
English

References (61)

Polyethylene glycol-coated biocompatible surfaces journal January 2000
Nitrilotriacetic Acid Functionalized Graft Copolymers: A Polymeric Interface for Selective and Reversible Binding of Histidine-Tagged Proteins journal January 2006
Optical and surface-analytical methods for the characterization of ultrathin organic films journal January 1991
Soft Lithographic Patterning of Hyaluronic Acid on Hydrophilic Substrates Using Molding and Printing journal April 2004
High molecular weight hydrophilic functional copolymers by free-radical copolymerization of acrylamide and of N -acryloylmorpholine with N -acryloxysuccinimide: Application to the synthesis of a graft copolymer: High MW Hydrophilic Functional Copolymers journal February 2003
Theoretical analysis of adsorption thermodynamics for charged peptide residues on SAM surfaces of varying functionality journal December 2002
SPR Studies of the Nonspecific Adsorption Kinetics of Human IgG and BSA on Gold Surfaces Modified by Self-Assembled Monolayers (SAMs) journal January 1997
Ni–NTA–Gold Clusters Target His-Tagged Proteins journal September 1999
Affinity purification of histidine-tagged proteins journal October 1993
Systems for the detection and analysis of protein–protein interactions journal June 2006
Biomedical surface science: Foundations to frontiers journal March 2002
The interaction of proteins and cells with self-assembled monolayers of alkanethiolates on gold and silver journal August 1999
Functionalized gold nanoparticles: Synthesis, structure and colloid stability journal March 2009
The interaction of proteins with solid surfaces journal February 2004
Resistance to Nonspecific Protein Adsorption by Poly(vinyl alcohol) Thin Films Adsorbed to a Poly(styrene) Support Matrix Studied Using Surface Plasmon Resonance journal September 2001
Fabrication of Histidine-Tagged Fusion Protein Arrays for Surface Plasmon Resonance Imaging Studies of Protein−Protein and Protein−DNA Interactions journal August 2003
Double-Hexahistidine Tag with High-Affinity Binding for Protein Immobilization, Purification, and Detection on Ni−Nitrilotriacetic Acid Surfaces journal April 2006
Nanoparticle-Enhanced Diffraction Gratings for Ultrasensitive Surface Plasmon Biosensing journal August 2007
Nitrilotriacetic Acid-Coated Magnetic Nanoparticles as Affinity Probes for Enrichment of Histidine-Tagged Proteins and Phosphorylated Peptides journal September 2007
XPS, TOF-SIMS, NEXAFS, and SPR Characterization of Nitrilotriacetic Acid-Terminated Self-Assembled Monolayers for Controllable Immobilization of Proteins journal April 2008
A Self-Assembled Monolayer for the Binding and Study of Histidine-Tagged Proteins by Surface Plasmon Resonance journal January 1996
Gold Nanoparticles in Biology: Beyond Toxicity to Cellular Imaging journal December 2008
Self-Assembled Monolayers and Polymer Brushes in Biotechnology:  Current Applications and Future Perspectives journal September 2005
Synthesis of Nickel−Nitrilotriacetic Acid Coupled Single-Walled Carbon Nanotubes for Directed Self-Assembly with Polyhistidine-Tagged Proteins journal February 2008
Self-Assembled Monolayers of Thiolates on Metals as a Form of Nanotechnology journal April 2005
Enzyme immobilization by condensation copolymerization into crosslinked polyacrylamide gels journal September 1980
Surface Adsorption and Transfer of Organomercaptans to Colloidal Gold and Direct Identification by Matrix Assisted Laser Desorption/Ionization Mass Spectrometry journal May 2004
Factors that Determine the Protein Resistance of Oligoether Self-Assembled Monolayers − Internal Hydrophilicity, Terminal Hydrophilicity, and Lateral Packing Density journal August 2003
Nanosensors Based on Responsive Polymer Brushes and Gold Nanoparticle Enhanced Transmission Surface Plasmon Resonance Spectroscopy journal December 2004
Kinase-Catalyzed Modification of Gold Nanoparticles:  A New Approach to Colorimetric Kinase Activity Screening journal January 2006
Quantitative Imaging of Protein Adsorption on Patterned Organic Thin-Film Arrays Using Secondary Electron Emission journal May 2006
Dual-Mode Fluorophore-Doped Nickel Nitrilotriacetic Acid-Modified Silica Nanoparticles Combine Histidine-Tagged Protein Purification with Site-Specific Fluorophore Labeling journal October 2007
Spontaneous Polymer Thin Film Assembly and Organization Using Mutually Immiscible Side Chains journal January 1996
Multifunctional Monolayer Assemblies for Reversible Direct Fluorescence Transduction of Protein−Ligand Interactions at Surfaces journal May 1999
Nanoscale Optical Biosensor:  Short Range Distance Dependence of the Localized Surface Plasmon Resonance of Noble Metal Nanoparticles journal June 2004
Anisotropic Metal Nanoparticles:  Synthesis, Assembly, and Optical Applications journal July 2005
Molecular Conformation and Solvation of Oligo(ethylene glycol)-Terminated Self-Assembled Monolayers and Their Resistance to Protein Adsorption journal November 1997
Size and Temperature Dependence of the Plasmon Absorption of Colloidal Gold Nanoparticles journal May 1999
Formation and Patterning of Self-Assembled Monolayers Derived from Long-Chain Organosilicon Amphiphiles and Their Use as Templates in Materials Microfabrication journal July 2000
Polymeric Thin Films That Resist the Adsorption of Proteins and the Adhesion of Bacteria journal February 2001
Assembly and Characterization of SAMs Formed by the Adsorption of Alkanethiols on Zinc Selenide Substrates journal May 2001
A Survey of Structure−Property Relationships of Surfaces that Resist the Adsorption of Protein journal September 2001
Functionalizing Surfaces with Nickel Ions for the Grafting of Proteins journal April 2003
Adsorption of Proteins to Hydrophobic Sites on Mixed Self-Assembled Monolayers journal March 2003
Self-Assembled Monolayers of Methyl 1-Thiahexa(ethylene oxide) for the Inhibition of Protein Adsorption journal May 2002
Self-Assembled Monolayers of an Oligo(ethylene oxide) Disulfide and Its Corresponding Thiol Assembled from Water:  Characterization and Protein Resistance journal January 2008
Self-Assembled Monolayers Containing Terminal Mono-, Bis-, and Tris-nitrilotriacetic Acid Groups:  Characterization and Application journal April 2008
A Highly Stable Nonbiofouling Surface with Well-Packed Grafted Zwitterionic Polysulfobetaine for Plasma Protein Repulsion journal April 2008
Quantitative Interpretation of the Response of Surface Plasmon Resonance Sensors to Adsorbed Films journal September 1998
Preparation of Mixed Self-Assembled Monolayers (SAMs) That Resist Adsorption of Proteins Using the Reaction of Amines with a SAM That Presents Interchain Carboxylic Anhydride Groups journal July 2000
Distance-Dependent Fluorescence Quenching on Gold Nanoparticles Ensheathed with Layer-by-Layer Assembled Polyelectrolytes journal March 2006
Textural guidance cues for controlling process outgrowth of mammalian neurons journal January 2009
Magnetic nanoparticle design for medical diagnosis and therapy journal January 2004
Tailoring plasmonic substrates for surface enhanced spectroscopies journal January 2008
Ni–nitrilotriacetic acid-modified quantum dots as a site-specific labeling agent of histidine-tagged proteins in live cells journal January 2008
A study of the nucleation and growth processes in the synthesis of colloidal gold journal January 1951
Optically tunable nanoparticle contrast agents for early cancer detection: model-based analysis of gold nanoshells journal January 2005
Molecular-Level Control over Surface Order in Self-Assembled Monolayer Films of Thiols on Gold journal April 1988
Using Self-Assembled Monolayers to Understand the Interactions of Man-made Surfaces with Proteins and Cells journal June 1996
Antibody-Antigen Complexes journal June 1990
Notizen: Radiative Decay of Non Radiative Surface Plasmons Excited by Light journal December 1968

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