Optical recognition of salivary proteins by use of molecularly imprinted poly(ethylene-co-vinyl alcohol)/quantum dot composite nanoparticles

Molecularly imprinted polymers (MIPs) have long been studied for applications in biomolecule recognition and binding; compared with natural antibodies, they may offer advantages in cost and stability. We report on the development of MIPs that “self-report” concentrations of bound analytes via fluorescence changes in embedded quantum dots (QDots). Composite QDot/MIPs were prepared using phase inversion of poly(ethylene-co-vinyl alcohol) (EVAL) solutions with various ethylene mole ratios in the presence of salivary target molecules (e.g. amylase, lipase, and lysozyme). These major protein components of saliva have been implicated as possible biomarkers for pancreatic cancer. The optimum (highest imprinting effectiveness) ethylene mole ratios of the commercially available EVALs were found to be 32, 38, and 44 mol% for the imprinting of amylase, lipase, and lysozyme, respectively. QD fluorescence quenching was observed on binding of analytes to composite MIPs in a concentration-dependent manner, and was used to construct calibration curves. Finally, the composite MIP particles were used for the quantitative detection of amylase, lipase, and lysozyme in real samples (saliva) and compared with a commercial Architect ci 8200 chemical analysis system.

[1]  Yunhua He,et al.  BSA-imprinted synthetic receptor for reversible template recognition. , 2009, Journal of separation science.

[2]  I. Luzinov,et al.  Toward protein imprinting with polymer brushes , 2009, Biointerphases.

[3]  T. Chou,et al.  Albumin molecularly imprinted polymer prepared with a semi-rigid crosslinker in mixed organic/aqueous media , 2009 .

[4]  Tse-Chuan Chou,et al.  The microcontact imprinting of proteins: the effect of cross-linking monomers for lysozyme, ribonuclease A and myoglobin. , 2006, Biosensors & bioelectronics.

[5]  J. Blumberg,et al.  Total Protein of Whole Saliva as a Biomarker of Anaerobic Threshold , 2009, Research quarterly for exercise and sport.

[6]  J. Goudswaard,et al.  Measurement of Salivary Lysozyme , 1978, Journal of dental research.

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

[8]  Xiwen He,et al.  Preparation of core-shell magnetic molecularly imprinted polymer nanoparticles for recognition of bovine hemoglobin. , 2009, Chemistry, an Asian journal.

[9]  Formation and recognition characteristics of albumin-imprinted poly(ethylene-co-vinyl-alcohol) membranes. , 2009, Journal of nanoscience and nanotechnology.

[10]  K. Lorentz.  Approved Recommendation on IFCC Methods for the Measurement of Catalytic Concentration of Enzymes Part 9. IFCC Method for α-Amylase (1,4-α-D-Glucan 4-Glucanohydrolase, EC 3.2.1.1) , 1998 .

[11]  Nicholas W Turner,et al.  From 3D to 2D: A Review of the Molecular Imprinting of Proteins , 2006, Biotechnology progress.

[12]  D. Guldi,et al.  Conjugating luminescent CdTe quantum dots with biomolecules. , 2008, The journal of physical chemistry. B.

[13]  L. Brus,et al.  Quantum size effects in the redox potentials, resonance Raman spectra, and electronic spectra of CdS crystallites in aqueous solution , 1983 .

[14]  Y. Tong,et al.  Preparation of superparamagnetic ribonuclease A surface-imprinted submicrometer particles for protein recognition in aqueous media. , 2007, Analytical chemistry.

[15]  Stellan Hjertén,et al.  Highly selective artificial gel antibodies for detection and quantification of biomarkers in clinical samples. II. Albumin in body fluids of patients with neurological disorders. , 2008, Journal of separation science.

[16]  T. Chou,et al.  Synthesis of and recognition by ribonuclease A imprinted polymers , 2006, Nanotechnology.

[17]  Mei-Hwa Lee,et al.  Instant formation of molecularly imprinted poly(ethylene-co-vinyl alcohol)/quantum dot composite nanoparticles and their use in one-pot urinalysis. , 2009, Biosensors & bioelectronics.

[18]  Adil Denizli,et al.  Protein recognition via ion-coordinated molecularly imprinted supermacroporous cryogels. , 2008, Journal of chromatography. A.

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

[20]  Yan Lu,et al.  Preparation and recognition of surface molecularly imprinted core–shell microbeads for protein in aqueous solutions , 2009 .

[21]  Meiping Zhao,et al.  Molecular imprinting of protein by coordinate interaction , 2009 .

[22]  L. Cheng,et al.  Phase Behavior of EVAL Polymers in Water-2-Propanol Cosolvent , 1998 .

[23]  Xiwen He,et al.  m-Aminophenylboronic Acid as a Functional Monomer for Fabricating Molecularly Imprinted Polymer for the Recognition of Bovine Serum Albumin. , 2008, Reactive & functional polymers.

[24]  D. Puleo,et al.  Protein Binding to Peptide-Imprinted Porous Silica Scaffolds. , 2008, Chemical engineering journal.

[25]  Ying Wang,et al.  Molecular imprinted polymer with positively charged assistant recognition polymer chains for adsorption/enrichment of low content target protein , 2008 .

[26]  Yizhe Jiang,et al.  Preparation of superparamagnetic ribonuclease A surface-imprinted submicrometer particles for protein recognition in aqueous media. , 2007 .

[27]  Xiwen He,et al.  Preparation of bovine hemoglobin-imprinted polymer beads via the photografting surface-modified method , 2008 .

[28]  U. Nater,et al.  Salivary alpha-amylase as a non-invasive biomarker for the sympathetic nervous system: Current state of research , 2009, Psychoneuroendocrinology.

[29]  Xiwen He,et al.  Optimum Conditions of Separation Selectivity Based on Molecularly Imprinted Polymers of Bovine Serum Albumin Formed on Surface of Aminosilica , 2009 .

[30]  T. Caykara,et al.  Dependence of protein recognition of temperature-sensitive imprinted hydrogels on preparation temperature. , 2009, Macromolecular bioscience.

[31]  Y. Sakai,et al.  Synthesis of polymer particles with specific lysozyme recognition sites by a molecular imprinting technique , 2001 .

[32]  T. Guo,et al.  Protein recognition onto silica particles using chitosan as intermedium substrate. , 2009, Journal of biomedical materials research. Part A.

[33]  Tse-Chuan Chou,et al.  Incorporation of styrene enhances recognition of ribonuclease A by molecularly imprinted polymers. , 2006, Biosensors & bioelectronics.

[34]  Hung-Yin Lin,et al.  Urinalysis with molecularly imprinted poly(ethylene-co-vinyl alcohol) potentiostat sensors. , 2009, Biosensors & bioelectronics.

[35]  Karsten Haupt,et al.  Polymer Films Composed of Surface‐Bound Nanofilaments with a High Aspect Ratio, Molecularly Imprinted with Small Molecules and Proteins , 2009 .

[36]  Nicolas Rohleder,et al.  Determinants of salivary α-amylase in humans and methodological considerations , 2009, Psychoneuroendocrinology.

[37]  A. Alivisatos.  Semiconductor Clusters, Nanocrystals, and Quantum Dots , 1996, Science.

[38]  G. Demirel,et al.  Hemoglobin recognition of molecularly imprinted hydrogels prepared at different pHs. , 2008, Analytica chimica acta.

[39]  Lei Qin,et al.  Surface-modified polystyrene beads as photografting imprinted polymer matrix for chromatographic separation of proteins. , 2009, Journal of chromatography. A.

[40]  Yanxiu Zhou,et al.  A potentiometric protein sensor built with surface molecular imprinting method. , 2008, Biosensors & bioelectronics.

[41]  Nicholas A. Peppas,et al.  Configurational Biomimetic Imprinting for Protein Recognition: Structural Characteristics of Recognitive Hydrogels , 2008 .

[42]  Junfu Wei,et al.  Preparation of bovine serum albumin‐imprinted calcium polyacrylate/alginate hybrid microspheres via Ca2+ crosslinking , 2009 .

[43]  Sergey Piletsky,et al.  Surface imprinted beads for the recognition of human serum albumin. , 2007, Biosensors & bioelectronics.

[44]  T. Chou,et al.  Albumin molecularly imprinted polymer with high template affinity — Prepared by systematic optimization in mixed organic/aqueous media , 2009 .

[45]  Wei Zhang,et al.  Novel surface modified molecularly imprinted polymer using acryloyl-beta-cyclodextrin and acrylamide as monomers for selective recognition of lysozyme in aqueous solution. , 2009, Journal of chromatography. A.