EL-Sharif, Hazim F., Hawkins, Daniel M., Stevenson, Derek and Reddy, Subrayal M (2014) Determination of protein binding affinities within hydrogel-based molecularly imprinted polymers (HydroMIPs). Physical Chemistry Chemical Physics, 16 (29). pp. 15483-15489. ISSN 1463-9076
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Official URL: http://dx.doi.org/10.1039/c4cp01798f
Hydrogel-based molecularly imprinted polymers (HydroMIPs) were prepared for several proteins (haemoglobin, myoglobin and catalase) using a family of acrylamide-based monomers. Protein aﬃnity towards the HydroMIPs was investigated under equilibrium conditions and over a range of concentrations using specific binding with Hill slope saturation profiles. We report HydroMIP binding aﬃnities, in terms of equilibrium dissociation constants (Kd) within the micro-molar range (25 ± 4 mM, 44 ± 3 mM, 17 ± 2 mM for haemoglobin, myoglobin and catalase respectively within a polyacrylamide-based MIP). The extent of non-specific binding or cross-selectivity for non-target proteins has also been assessed. It is concluded that both selectivity and aﬃnity for both cognate and non-cognate proteins towards the MIPs were dependent on the concentration and the complementarity of their structures and size. This is tentatively attributed to the formation of protein complexes during both the polymerisation and rebinding stages at high protein concentrations. We have used atomic force spectroscopy to characterize molecular interactions in the MIP cavities using protein-modified AFM tips. Attractive and repulsive force curves were obtained for the MIP and NIP (non-imprinted polymer) surfaces (under protein loaded or unloaded states). Our force data suggest that we have produced selective cavities for the template protein in the MIPs and we have been able to quantify the extent of non-specific protein binding on, for example, a non-imprinted polymer (NIP) control surface.
|Subjects:||Physical sciences > Chemistry|
|Schools:||Faculty of Science and Technology > School of Physical Sciences and Computing|
|Deposited By:||Nuala Jones|
|Deposited On:||16 Feb 2016 17:55|
|Last Modified:||24 Oct 2016 05:05|
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