In the protein adsorption experiments,hen egg white lysozyme
and BSA whose molecular weights are 14 kDa and 66k Da,
respectively, were used a smodel proteins.Lysozyme or BSA was dissolved
in 7m L of 20 mM phosphate-bufferedsaline(PBS) solution(pH7.4)
and 35 mg of the hydrogel was added to the protein-containing
buffer. The adsorption experiments were then conducted in a
the rmostat-containing water bathat 25 ◦C for 24 h,with
shaking at arate of 120 rpm. At the end of the binding period,the
mixture was centrifuged at 15,000 rpm for 2 min, and the
protein adsorbed hydroge lwas obtained (Kono, Oeda,&Nakamura,2013).
The proteinconcentrationofthesupernatantwasestimatedata
wavelength of280nmusinganEvolution201UV–visspectropho-
tometer. Theamountofproteinadsorbedtothehydrogelswas
determined bythefollowingequation:
mixture wascentrifugedat15,000rpmfor2min,andtheprotein-
adsorbed hydrogelwasobtained(Kono, Oeda,&Nakamura,2013).
The proteinconcentrationofthesupernatantwasestimatedata
wavelength of280nmusinganEvolution201UV–visspectropho-
tometer. Theamountofproteinadsorbedtothehydrogelswas
determined bythefollowingequation: