Sensing HIV related protein using epitope imprinted hydrophilic polymer coated quartz crystal microbalance.

We have developed a biomimetic sensor for the detection of human immunodeficiency virus type 1 (HIV-1) related protein (glycoprotein 41, gp41) based on epitope imprinting technique. gp41 is the transmembrane protein of HIV-1 and plays an important role in membrane fusion between viruses and infected cells. It is an important index for determining the extent of HIV-1 disease progression and the efficacy of therapeutic intervention. In this work, dopamine was used as the functional monomer and polymerized on the surface of quartz crystal microbalance (QCM) chip in the presence of template, a synthetic peptide with 35 amino acid residues, analogous to residues 579-613 of the gp41. This process resulted in grafting a hydrophilic molecularly imprinted polymer (MIP) film on the QCM chip. QCM measurement showed that the resulting MIP film not only had a great affinity towards the template peptide, but also could bind the corresponding gp41 protein specifically. The dissociation constant (K(d)) of MIP for the template peptide was calculated to be 3.17 nM through Scatchard analysis, which was similar to those of monoclonal antibodies. Direct detection of the gp41 was achieved quantitatively using the resulting MIP-based biomimetic sensor. The detection limit of gp41 was 2 ng/mL, which was comparable to the reported ELISA method. In addition, the practical analytical performance of the sensor was examined by evaluating the detection of gp41 in human urine samples with satisfactory results.

[1]  J. González-Ros,et al.  Structure and interaction with membrane model systems of a peptide derived from the major epitope region of HIV protein gp41: implications on viral fusion mechanism. , 2001, Biochemistry.

[2]  Yu Hoshino,et al.  Synthetic polymer nanoparticles with antibody-like affinity for a hydrophilic peptide. , 2010, ACS nano.

[3]  Itamar Willner,et al.  Imprinting of molecular recognition sites through electropolymerization of functionalized Au nanoparticles: development of an electrochemical TNT sensor based on pi-donor-acceptor interactions. , 2008, Journal of the American Chemical Society.

[4]  I. Willner,et al.  Ultrasensitive surface plasmon resonance detection of trinitrotoluene by a bis-aniline-cross-linked Au nanoparticles composite. , 2009, Journal of the American Chemical Society.

[5]  A. Rachkov,et al.  Recognition of oxytocin and oxytocin-related peptides in aqueous media using a molecularly imprinted polymer synthesized by the epitope approach. , 2000, Journal of chromatography. A.

[6]  A. Turner,et al.  Molecularly imprinted polymers for the recognition of proteins: the state of the art. , 2007, Biosensors & bioelectronics.

[7]  Shubhra Gangopadhyay,et al.  Detection of nitroaromatic explosives using a fluorescent-labeled imprinted polymer. , 2010, Analytical chemistry.

[8]  M. V. D. Leeden.  Are conformational changes, induced by osmotic pressure variations, the underlying mechanism of controlling the adhesive activity of mussel adhesive proteins? , 2005 .

[9]  G. Wulff.  Molecular Imprinting in Cross‐Linked Materials with the Aid of Molecular Templates— A Way towards Artificial Antibodies , 1995 .

[10]  Lei Ye,et al.  Molecular imprinting: Synthetic materials as substitutes for biological antibodies and receptors , 2008 .

[11]  A. Turner,et al.  Surface-grafted molecularly imprinted polymers for protein recognition. , 2001, Analytical chemistry.

[12]  A. Turner,et al.  Molecularly imprinted sorbent assays: recent developments and applications. , 2009, Chemistry.

[13]  Karsten Haupt,et al.  Molecularly imprinted microgels as enzyme inhibitors. , 2009, Journal of the American Chemical Society.

[14]  T. Matthews,et al.  The hydrophobic pocket contributes to the structural stability of the N-terminal coiled coil of HIV gp41 but is not required for six-helix bundle formation. , 2003, Biochemistry.

[15]  S. Munir Alam,et al.  Human Immunodeficiency Virus Type 1 gp41 Antibodies That Mask Membrane Proximal Region Epitopes: Antibody Binding Kinetics, Induction, and Potential for Regulation in Acute Infection , 2007, Journal of Virology.

[16]  P S Kim,et al.  Evidence that a prominent cavity in the coiled coil of HIV type 1 gp41 is an attractive drug target. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[17]  J. Waite,et al.  Cross-linking in adhesive quinoproteins: studies with model decapeptides. , 2000, Biochemistry.

[18]  Y. Okahata,et al.  Peptide imprinted polymer nanoparticles: a plastic antibody. , 2008, Journal of the American Chemical Society.

[19]  Zilun Chen,et al.  Imprinting of protein over silica nanoparticles via surface graft copolymerization using low monomer concentration. , 2010, Biosensors & bioelectronics.

[20]  Jeffrey H. Chuang,et al.  A molecular-imprint nanosensor for ultrasensitive detection of proteins. , 2010, Nature nanotechnology.

[21]  M. Mizuhata,et al.  Protein-templated organic/inorganic hybrid materials prepared by liquid-phase deposition. , 2007, Journal of the American Chemical Society.

[22]  Xiu‐Ping Yan,et al.  Discrimination of saccharides with a fluorescent molecular imprinting sensor array based on phenylboronic acid functionalized mesoporous silica. , 2009, Analytical chemistry.

[23]  John O'Mahony,et al.  Molecular imprinting science and technology: a survey of the literature for the years up to and including 2003 , 2006, Journal of molecular recognition : JMR.

[24]  Wayne C Koff,et al.  HIV vaccine design and the neutralizing antibody problem , 2004, Nature Immunology.

[25]  Huang-Hao Yang,et al.  Template synthesized molecularly imprinted polymer nanotube membranes for chemical separations. , 2006, Journal of the American Chemical Society.

[26]  Tzong-Zeng Wu,et al.  Recognition of dengue virus protein using epitope-mediated molecularly imprinted film. , 2005, Analytical chemistry.

[27]  J. Cladera,et al.  Effect of cholesterol on the interaction of the HIV GP41 fusion peptide with model membranes. Importance of the membrane dipole potential. , 2006, Biochemistry.

[28]  D. Tai,et al.  Epitope-cavities generated by molecularly imprinted films measure the coincident response to anthrax protective antigen and its segments. , 2010, Analytical chemistry.

[29]  Yong Li,et al.  Protein recognition via surface molecularly imprinted polymer nanowires. , 2006, Analytical chemistry.

[30]  H. Ju,et al.  A Molecularly Imprinted Copolymer Designed for Enantioselective Recognition of Glutamic Acid , 2007 .

[31]  Karsten Haupt,et al.  Molecularly imprinted polymers: the next generation. , 2003, Analytical chemistry.

[32]  H. Katinger,et al.  Characterization of a trimeric MPER containing HIV-1 gp41 antigen. , 2009, Virology.

[33]  Anthony Turner,et al.  Too large to fit? Recent developments in macromolecular imprinting. , 2008, Trends in biotechnology.

[34]  Huangxian Ju,et al.  Surface molecularly imprinted nanowire for protein specific recognition. , 2008, Chemical communications.

[35]  Haeshin Lee,et al.  Mussel-Inspired Surface Chemistry for Multifunctional Coatings , 2007, Science.

[36]  H. Ju,et al.  Artificial receptor-functionalized nanoshell: facile preparation, fast separation and specific protein recognition , 2010, Nanotechnology.

[37]  Daming Gao,et al.  A surface functional monomer-directing strategy for highly dense imprinting of TNT at surface of silica nanoparticles. , 2007, Journal of the American Chemical Society.

[38]  K. Haupt,et al.  Toward the use of a molecularly imprinted polymer in doping analysis: selective preconcentration and analysis of testosterone and epitestosterone in human urine. , 2010, Analytical chemistry.

[39]  Wei Zhang,et al.  Composite of CdTe quantum dots and molecularly imprinted polymer as a sensing material for cytochrome c. , 2011, Biosensors & bioelectronics.

[40]  Klaus Mosbach,et al.  Drug assay using antibody mimics made by molecular imprinting , 1993, Nature.

[41]  K. Shea,et al.  Selective protein capture by epitope imprinting. , 2006, Angewandte Chemie.

[42]  Jianping Li,et al.  Fabrication of an oxytetracycline molecular-imprinted sensor based on the competition reaction via a GOD-enzymatic amplifier. , 2011, Biosensors & bioelectronics.

[43]  Rigoberto Advincula,et al.  Electropolymerization molecularly imprinted polymer (E-MIP) SPR sensing of drug molecules: pre-polymerization complexed terthiophene and carbazole electroactive monomers. , 2011, Biosensors & bioelectronics.

[44]  B. Sellergren,et al.  High-capacity hierarchically imprinted polymer beads for protein recognition and capture. , 2011, Angewandte Chemie.

[45]  Buddy D. Ratner,et al.  Template-imprinted nanostructured surfaces for protein recognition , 1999, Nature.

[46]  Huang-Hao Yang,et al.  Mussel-inspired molecularly imprinted polymer coating superparamagnetic nanoparticles for protein recognition , 2010 .