Abstract
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.
References
Lorentz K (2005) 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). Clin Chem Lab Med 36:185–203
Nater UM, Rohleder N (2009) Salivary alpha-amylase as a non-invasive biomarker for the sympathetic nervous system: current state of research. Psychoneuroendocrinology 34:486–496
Bortolini MJS, De Agostini GG, Reis IT, Lamounier RPMS, Blumberg JB, Espindola FS (2009) Total protein of whole saliva as a biomarker of anaerobic threshold. Res Q Exerc Sport 80:604–610
Rohleder N, Nater UM (2009) Determinants of salivary [alpha]-amylase in humans and methodological considerations. Psychoneuroendocrinology 34:469–485
Rossetti R, Nakahara S, Brus LE (1983) Quantum size effects in the redox potentials, resonance Raman spectra, and electronic spectra of CdS crystallites in aqueous solution. J Chem Phys 79:1086–1088
Alivisatos AP (1996) Semiconductor clusters, nanocrystals, and quantum dots. Science 271:933–937
Ye L, Mosbach K (2008) Molecular imprinting: synthetic materials as substitutes for biological antibodies and receptors. Chem Mater 20:859–868
Bossi A, Bonini F, Turner APF, Piletsky SA (2007) Molecularly imprinted polymers for the recognition of proteins: the state of the art. Biosens Bioelectron 22:1131–1137
Turner NW, Jeans CW, Brain KR, Allender CJ, Hlady V, Britt DW (2006) From 3D to 2D: a review of the molecular imprinting of proteins. Biotechnol Prog 22:1474–1489
Bonini F, Piletsky S, Turner APF, Speghini A, Bossi A (2007) Surface imprinted beads for the recognition of human serum albumin. Biosens Bioelectron 22:2322–2328
Zhou X, He X-W, Chen L-X, Li W-Y, Zhang Y-K (2009) Optimum conditions of separation selectivity based on molecularly imprinted polymers of bovine serum albumin formed on surface of Aminosilica. Chinese J Anal Chem 37:174–180
Zhao K, JWei u, Cheng G, Yang C, Chen L (2009) Preparation of bovine serum albumin-imprinted calcium polyacrylate alginate hybrid microspheres via Ca2+ crosslinking. J Appl Polym Sci 113:1133–1140
Zdyrko B, Hoy O, Luzinov I (2009) Toward protein imprinting with polymer brushes. Biointerphases 4:FA17–FA21
Wang J, Hua ZD, Chen ZY, Li YZ, Zhao MP (2009) Molecular imprinting of protein by coordinate interaction. Chin Chem Lett 20:747–750
Wang H, Li W, He X, Chen L, Zhang Y (2008) m-Aminophenylboronic acid as a functional monomer for fabricating molecularly imprinted polymer for the recognition of bovine serum albumin. React Funct Polym 68:1291–1296
Wang H, He Y, He X, Li W, Chen L, Zhang Y (2009) BSA-imprinted synthetic receptor for reversible template recognition. J Sep Sci 32:1981–1986
Lu Y, Yan C-L, Gao S-Y (2009) Preparation and recognition of surface molecularly imprinted core-shell microbeads for protein in aqueous solutions. Appl Surf Sci 255:6061–6066
Long Y, Sun Y, Wang Y, Xing X, Zhao Z, Wang C, Fan Y, Mi H (2008) Molecular imprinted polymer with positively charged assistant recognition polymer chains for adsorption/enrichment of low content target protein. Chin Sci Bull 53:2617–2623
Lee M-H, Thomas JL, Tasi S-B, Liu B-D, Lin H-Y (2009) Formation and Recognition Characteristics of Albumin-Imprinted Poly(Ethylene-co-Vinyl-Alcohol) Membranes. J Nanosci Nanotechnol 9:3469–3477
Hu C-H, Chou T-C (2009) Albumin molecularly imprinted polymer prepared with a semi-rigid crosslinker in mixed organic/aqueous media. Microchim Acta 165:399–405
Hu C-H, Chou T-C (2009) Albumin molecularly imprinted polymer with high template affinity – Prepared by systematic optimization in mixed organic/aqueous media. Microchem J 91:53–58
Ghasemzadeh N, Nyberg F, Hjertén S (2008) Highly selective artificial gel antibodies for detection and quantification of biomarkers in clinical samples. II. Albumin in body fluids of patients with neurological disorders. J Sep Sci 31:3954–3958
Brown ME, Puleo DA (2008) Protein binding to peptide-imprinted porous silica scaffolds. Chem Eng J 137:97–101
Zhang W, Qin L, He X-W, Li W-Y, Zhang Y-K (2009) Novel surface modified molecularly imprinted polymer using acryloyl-[beta]-cyclodextrin and acrylamide as monomers for selective recognition of lysozyme in aqueous solution. J Chromatogr 1216:4560–4567
Qin L, He X-W, Zhang W, Li W-Y, Zhang Y-K (2009) Surface-modified polystyrene beads as photografting imprinted polymer matrix for chromatographic separation of proteins. J Chromatogr 1216:807–814
Huang C-Y, Tsai T-C, Thomas JL, Lee M-H, Liu B-D, Lin H-Y (2009) Urinalysis with molecularly imprinted poly(ethylene-co-vinyl alcohol) potentiostat sensors. Biosens Bioelectron 24:2611–2617
Bergmann NM, Peppas NA (2008) Configurational biomimetic imprinting for protein recognition: structural characteristics of recognitive hydrogels. Ind Eng Chem Res 47:9099–9107
Bereli N, Me A, Baydemir G, Say R, Galaev IY, Denizli A (2008) Protein recognition via ion-coordinated molecularly imprinted supermacroporous cryogels. J Chromatogr 1190:18–26
Lin H-Y, Hsu C-Y, Thomas JL, Wang S-E, Chen H-C, Chou T-C (2006) The microcontact imprinting of proteins: The effect of cross-linking monomers for lysozyme, ribonuclease A and myoglobin. Biosens Bioelectron 22:534–543
Hirayama K, Sakai Y, Kameoka K (2001) Synthesis of polymer particles with specific lysozyme recognition sites by a molecular imprinting technique. J Appl Polym Sci 81:3378–3387
Tan CJ, Tong YW (2006) Preparation of superparamagnetic ribonuclease a surface-imprinted submicrometer particles for protein recognition in Aqueous Media. Anal Chem 79:299–306
Hsu C-Y, Lin H-Y, Thomas JL, Wu B-T, Chou T-C (2006) Incorporation of styrene enhances recognition of ribonuclease A by molecularly imprinted polymers. Biosens Bioelectron 22:355–363
Hsu C-Y, Lin H-Y, Thomas JL, Chou T-C (2006) Synthesis of and recognition by ribonuclease A imprinted polymers. Nanotechnology 17:S77–S83
Xia Y-Q, Guo T-Y, Zhao H-L, Song M-D, Zhang B-H, Zhang B-L (2009) Protein recognition onto silica particles using chitosan as intermedium substrate. J Biomed Materi Res 90A:326–332
Wang Y, Zhou Y, Sokolov J, Rigas B, Levon K, Rafailovich M (2008) A potentiometric protein sensor built with surface molecular imprinting method. Biosens Bioelectron 24:162–166
Uysal A, Demirel G, Turan E, Çaykara T (2008) Hemoglobin recognition of molecularly imprinted hydrogels prepared at different pHs. Anal Chim Acta 625:110–115
Gai Q, Liu Q, Li W, He X, Chen L, Zhang Y (2008) Preparation of bovine hemoglobin-imprinted polymer beads via the photografting surface-modified method. Front Chem Chin 3:370–377
Li L, He X, Chen L, Zhang Y (2009) Preparation of core-shell magnetic molecularly imprinted polymer nanoparticles for recognition of bovine hemoglobin. Chem Asian J 4:286–293
Linares AV, Vandevelde F, Pantigny J, Falcimaigne-Cordin A, Haupt K (2009) Polymer films composed of surface-bound nanofilaments with a high aspect ratio, molecularly imprinted with small molecules and proteins. Adv Funct Mater 19:1299–1303
Turan E, Özçetin G, Caykara T (2009) Dependence of protein recognition of temperature-sensitive imprinted hydrogels on preparation temperature. Macromol Biosci 9:421–428
Lin H-Y, Ho M-S, Lee M-H (2009) Instant formation of molecularly imprinted poly(ethylene-co-vinyl alcohol)/quantum dot composite nanoparticles and their use in one-pot urinalysis. Biosens Bioelectron 25:579–586
Young TH, Cheng LP, Hsieh CC, Chen LW (1998) Phase behavior of EVAL polymers in water-2-Propanol Cosolvent. Macromolecules 31:1229–1235
Gerhards C, Schulz-Drost C, Sgobba V, Guldi DM (2008) Conjugating Luminescent CdTe Quantum Dots with Biomolecules†. J Phys Chem B 112:14482–14491
Virella G, Goudswaard J (1978) Measurement of salivary lysozyme. J Dent Res 57:326–328
Acknowledgements
We appreciate financial support from National Science Council of ROC under Contract No. NSC 98-2220-E-390-002.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Lee, MH., Chen, YC., Ho, MH. et al. Optical recognition of salivary proteins by use of molecularly imprinted poly(ethylene-co-vinyl alcohol)/quantum dot composite nanoparticles. Anal Bioanal Chem 397, 1457–1466 (2010). https://doi.org/10.1007/s00216-010-3631-x
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00216-010-3631-x