Figure 4.

Biophysical characterization indicates copolymer-assisted refolding. a) Circular dichroism wavelength scans of HRP (dashed lines) and HRP-EP1 (solid lines) at room temperature (black), upon heating (red), and after cooling for 24hrs (blue), demonstrating that HRP-EP1 promotes retention of secondary structure in HRP during thermal stress and promotes significant protein refolding in comparison to HRP control. b) Pair-distance distribution function of HRP and HRP-EP1 by small-angle X-ray scattering demonstrating retained HRP-PPH morphology and size after exposure to thermal stress in comparison to native enzyme. c) Guinier analysis of HRP and HRP-EP1 before and after heating suggesting the development of a denatured or aggregated sub-population of HRP (blue line) in comparison to a single species observed in HRP, HRP-EP1, and HRP-EP1 after thermal stress (red lines). d) Dynamic light scattering size distributions of HRP with and without polymer EP1, demonstrating that no larger structures were observed after mixing. e) Surface thickness measured by Quartz crystal microbalance with dissipation after direct adsorption of HRP (t = 22 min) followed by injection of polymer EP1 (t = 82 min).