Abstract
Sol–gels are seeing widespread interest as suitable materials for the immobilization of biomolecules in applications ranging from optical coatings to specialty biocatalysts. Although there are numerous studies that have characterized these materials in terms of their macroscopic properties, few studies have examined and correlated these properties at the microscopic level. This study describes a spin-coating technique for the preparation of aluminum-supported sol–gel thin films containing immobilized lysozyme [E.C. 3.2.1.17] that are suitable for chemical mapping using FTIR microscopy operating in reflectance mode. This type of information can then be used to understand a variety of aspects of these materials which can be used for optimal engineering of these materials, as well as insightful design and modeling. A data analysis method was developed to extract information on chemical speciation and domain information on the materials from FTIR data matrices. Results from these studies indicated that, contrary to what might be expected, these sol–gels are not homogeneous on the microscopic level. Instead, they are heterogeneous in both the distribution of lysozyme and hydrophobic monomers at the scale investigated (20 μm resolution). The method described here has promise in terms of providing a non-invasive approach of chemically mapping concentrations of proteinaceous and related substances as well as chemical domains in situ in sol–gel thin films.
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References
Drauz K, Waldmann H (2002) Enzyme catalysis in organic synthesis. Wiley-VCH, Weinheim
Clark DS (1994) Trends Biotechnol 12:439. doi:10.1016/0167-7799(94)90018-3
Jürgen-Lohmann DL, Legge RL (2006) Enzyme Microb Technol 39:626. doi:10.1016/j.enzmictec.2005.11.015
Gill I, Ballesteros A (2000) Trends Biotechnol 18:282. doi:10.1016/S0167-7799(00)01457-8
Jin W, Brennan JD (2002) Anal Chim Acta 461:1. doi:10.1016/S0003-2670(02)00229-5
Pierre AC (2004) Biocat Biotransf 22:145. doi:10.1080/10242420412331283314
Günzler H, Gremlich HU (2002) IR spectroscopy. Wiley-VCH, Weinheim
Gremlich HU, Yan B (2001) Infrared and Raman spectroscopy of biological materials. Marcel Dekker, New York
Schultz CP (2001) Spectroscopy 16:24
Wetzel DL, LeVine SM (1999) Science 285:1225. doi:10.1126/science.285.5431.1224
Bhargava R, Wall BG, Koenig JL (2000) Appl Spectrosc 54:470. doi:10.1366/0003702001949870
Bhargava R, Wang SQ, Koenig JL (2003) Adv Polym Sci 163:137
Sammon C, Boussetta S, Melia C (2002) Macromol Symp 184:357. doi:10.1002/1521-3900(200208)184:1<357::AID-MASY357>3.0.CO;2-5
Mei Y, Miller L, Gao W, Gross RA (2003) Biomacromolecules 4:70. doi:10.1021/bm025611t
Kistler SF, Schweizer PM (1997) Liquid film coating. Chapman & Hall, London
Huang YY, Chou KS (2003) Ceram Int 29:485. doi:10.1016/S0272-8842(02)00191-8
Taylor JF (2001) Met Finish 99:16. doi:10.1016/S0026-0576(01)80527-4
Clifford JS, Legge RL (2005) Biotechnol Bioeng 92:231. doi:10.1002/bit.20595
Wetzel DL, Striova J, Higgins DA, Collinson MM (2004) Vib Spectrosc 35:153. doi:10.1016/j.vibspec.2003.12.018
Higgins DA, Collinson MM (2005) Langmuir 21:9023. doi:10.1021/la050384c
Durrani CM, Donald AM (1995) Carbohydr Polym 28:297. doi:10.1016/0144-8617(95)00107-7
Acknowledgements
The authors are grateful to Dr. James Forrest and James Chang from the Polymer Physics group at the University of Waterloo, Waterloo, ON for sharing their spin coating equipment and expertise. RLL and LCS are grateful for funding from NSERC.
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Jürgen-Lohmann, D.L., Nacke, C., Legge, R.L. et al. Preparation and methodology for chemical mapping of sol–gel thin films containing lysozyme. J Sol-Gel Sci Technol 50, 77–86 (2009). https://doi.org/10.1007/s10971-009-1893-6
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DOI: https://doi.org/10.1007/s10971-009-1893-6