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BY-NC-ND 3.0 license Open Access Published by De Gruyter Open Access March 16, 2016

Development of protein-recognition SPR devices by combination of SI-ATRP with biomolecular imprinting using protein ligands

  • Rinyarat Naraprawatphong , Genta Kawanaka , Masayoshi Hayashi , Akifumi Kawamura and Takashi Miyata
From the journal Molecular Imprinting


Molecularly imprinted polymer brush layers and gel layers with both a lectin (ConA) and an antibody-IgG as biomolecular ligands for a target protein were formed on surface plasmon resonance (SPR) sensor chips via surface-initiated atom transfer radical polymerization (SIATRP) without and with a crosslinker, respectively. While the IgG-imprinted brush layers chip had almost the same affinity constant for target IgG as the nonimprinted brush layer chip, the affinity constant of the IgG-imprinted gel layer chip was approximately twice than that of the nonimprinted gel layer chip. These indicate that chemical crosslinks are very important factor to create distinct molecular recognition sites by molecular imprinting. Thus, biomolecular imprinting that uses biomolecular ligands and crosslinkers enables us to design polymer layer chips with distinct molecular recognition sites with a strong affinity for a target biomolecule. The molecularly imprinted gel layers chips with lectin and antibody ligands are promising candidates for fabricating SPR sensor systems to monitor target biomolecules such as proteins.


[1] Bier, F.F., F. Kleinjung, and F.W. Scheller, Real-time measurement of nucleic-acid hybridization using evanescentwave sensors: steps towards the genosensor. Sens. Actuators, B: Chemical, 1997. 38(1-3): p. 78-82. Search in Google Scholar

[2] Elkind, J.L., et al., Integrated analytical sensors: the use of the TISPR-1 as a biosensor. Sens. Actuators, B: Chemical, 1999. 54(1–2): p. 182-190. Search in Google Scholar

[3] Homola, J., Surface plasmon resonance sensors for detection of chemical and biological species. Chem. Rev., 2008. 108(2): p. 462-93. Search in Google Scholar

[4] Kooyman, R.P.H., et al., Surface plasmon resonance immunosensors: sensitivity considerations. Anal. Chim. Acta, 1988. 213: p. 35-45. Search in Google Scholar

[5] Liedberg, B., C. Nylander, and I. Lunström, Surface plasmon resonance for gas detection and biosensing. Sens. Actuators, 1983. 4: p. 299-304. 10.1016/0250-6874(83)85036-7Search in Google Scholar

[6] Schuster, S.C., et al., Assembly and function of a quaternary signal transduction complex monitored by surface plasmon resonance. Nature, 1993. 365(6444): p. 343-7. 10.1038/365343a0Search in Google Scholar

[7] Silin, V. and A. Plant, Biotechnological applications of surface plasmon resonance. Trends Biotechnol., 1997. 15(9): p. 353-359. 10.1016/S0167-7799(97)01085-8Search in Google Scholar

[8] Silin, V., H. Weetall, and D.J. Vanderah, SPR Studies of the Nonspecific Adsorption Kinetics of Human IgG and BSA on Gold Surfaces Modified by Self-Assembled Monolayers (SAMs). J. Colloid Interface Sci., 1997. 185(1): p. 94-103. 10.1006/jcis.1996.4586Search in Google Scholar

[9] Stojanovic, I., R.B. Schasfoort, and L.W. Terstappen, Analysis of cell surface antigens by Surface Plasmon Resonance imaging. Biosens. Bioelectron., 2014. 52: p. 36-43. 10.1016/j.bios.2013.08.027Search in Google Scholar

[10] Somasundaram, B., et al., Development of a surface plasmon resonance assay to measure the binding affinity of wild-type influenza neuraminidase and its H274Y mutant to the antiviral drug zanamivir. J. Mol. Recognit., 2015. 28(2): p. 87-95. 10.1002/jmr.2417Search in Google Scholar

[11] Catimel, B., et al., Kinetic analysis of the interaction between the monoclonal antibody A33 and its colonic epithelial antigen by the use of an optical biosensor. J. Chromatogr. A, 1997. 776(1): p. 15-30. 10.1016/S0021-9673(97)00087-3Search in Google Scholar

[12] Frederix, F., et al., Reduced nonspecific adsorption on covalently immobilized protein surfaces using poly(ethylene oxide) containing blocking agents. J. Biochem. Bioph. Methods, 2004. 58(1): p. 67-74. Search in Google Scholar

[13] Kuriu, Y., et al., SPR Signals of Three-dimensional Antibodyimmobilized Gel Layers Formed on Sensor Chips by Atom Transfer Radical Polymerization. Chem. Lett., 2012. 41(12): p. 1660-1662. Search in Google Scholar

[14] Ahmed, E.M., Hydrogel: Preparation, characterization, and applications: A review. J. Adv. Res., 2015. 6(2): p. 105-21. Search in Google Scholar

[15] Miyata, T., Preparation of smart soft materials using molecular complexes. Polymer Journal, 2010. 42(4): p. 277-289. 10.1038/pj.2010.12Search in Google Scholar

[16] Miyata, T., et al., Controlled permeation of model drugs through a bioconjugated membrane with antigen–antibody complexes as reversible crosslinks. Polym. J., 2010. 42(10): p. 834-837. Search in Google Scholar

[17] Miyata, T., N. Asami, and T. Uragami, A reversibly antigenresponsive hydrogel. Nature, 1999. 399(6738): p. 766-9. 10.1038/21619Search in Google Scholar PubMed

[18] Miyata, T., et al., Responsive behavior of tumor-markerimprinted hydrogels using macromolecular cross-linkers. J. Mol. Recognit., 2012. 25(6): p. 336-43. 10.1002/jmr.2190Search in Google Scholar PubMed

[19] Miyata, T., et al., Tumor marker-responsive behavior of gels prepared by biomolecular imprinting. Proc. Natl. Acad. Sci. U S A, 2006. 103(5): p. 1190-3. 10.1073/pnas.0506786103Search in Google Scholar PubMed PubMed Central

[20] Miyata, T., T. Uragami, and K. Nakamae, Biomolecule-sensitive hydrogels. Adv. Drug Deliv. Rev., 2002. 54(1): p. 79-98. Search in Google Scholar

[21] Mosbach, K., Molecular Imprinting. Trends Biochem.Sci, 1994. 19(1): p. 9-14. Search in Google Scholar

[22] Wulff, G., Molecular Imprinting in Cross-Linked Materials with the Aid of Molecular Templates— A Way towards Artificial Antibodies. Angew. Chem. Int. Ed. Engl, 1995. 34(17): p.1812-1832. Search in Google Scholar

[23] Shea, K., Molecular imprinting of synthetic network polymers: the de novo synthesis of macromolecular binding and catalytic sites. Trends Polym. Sci, 1994. 2(5):p. 155-173. Search in Google Scholar

[24] Vasapollo, G., et al., Molecularly imprinted polymers: present and future prospective. Int J Mol Sci, 2011. 12(9): p. 5908-45. 10.3390/ijms12095908Search in Google Scholar PubMed PubMed Central

[25] Byrne, M.E. and V. Salian, Molecular imprinting within hydrogels II: progress and analysis of the field. Int. J. Pharm., 2008. 364(2): p. 188-212. 10.1016/j.ijpharm.2008.09.002Search in Google Scholar PubMed

[26] Lv, Y., T. Tan, and F. Svec, Molecular imprinting of proteins in polymers attached to the surface of nanomaterials for selective recognition of biomacromolecules. Biotechnol. Adv., 2013. 31: p. 1172-86. Search in Google Scholar

[27] Takeuchi, T. and T. Hishiya, Molecular imprinting of proteins emerging as a tool for protein recognition. Org. Biomol. Chem., 2008. 6(14): p. 2459-67. Search in Google Scholar

[28] Bossi, A., et al., Molecularly imprinted polymers for the recognition of proteins: the state of the art. Biosens. Bioelectron., 2007. 22(6): p. 1131-7. 10.1016/j.bios.2006.06.023Search in Google Scholar PubMed

[29. Lépinay, S., Kham, K., Millot, M-C., Carbonnier, B., In-situ polymerized molecularly imprinted polymeric thin films used as sensing layers in surface plasmon resonance sensors : Mini-review focused on 2010–2011.Chem. Pap., 2012, 66(5): p. 340-351. 10.2478/s11696-012-0134-6Search in Google Scholar

[30] Morelli, I., et al., Molecularly imprinted submicronspheres for applications in a novel model biosensor-film. Sens. Actuators, B: Chemical, 2010. 150(1): p. 394-401. 10.1016/j.snb.2010.06.046Search in Google Scholar

[31] Barbey, R., et al., Polymer brushes via surface-initiated controlled radical polymerization: synthesis, characterization, properties, and applications. Chem. Rev., 2009. 109(11): p. 5437-527. Search in Google Scholar

[32] Miyata, T., et al., Structural design of stimuli-responsive bioconjugated hydrogels that respond to a target antigen. J. Polym. Sci., Part B: Polym. Phys, 2009. 47(21): p. 2144-2157. Search in Google Scholar

[33] Shoemaker, S. G., et al., Synthesis and properties of vinyl monomer/enzyme conjugates; Conjugation of L-Asparaginase with N-succinimidyl acrylate. Appl. Biochem. Bioethanol, 1987. 15: p. 11-24. Search in Google Scholar

[34] Kuriu, Y., et al., Formation of Thin Molecularly Imprinted Hydrogel Layers with Lectin Recognition Sites on SPR Sensor Chips by Atom Transfer Radical Polymerization. Chem. Lett., 2014. 43(6): p. 825-827. 10.1246/cl.140103Search in Google Scholar

Received: 2015-10-5
Accepted: 2016-2-10
Published Online: 2016-3-16

© 2016 Rinyarat Naraprawatphong et al.

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

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