Premium
Measuring the kinetics and activity of adsorbed proteins: In vitro lysozyme deposited onto hydrogel contact lenses over short time periods
Author(s) -
Hall B.,
Jones L.,
Forrest J. A.
Publication year - 2013
Publication title -
journal of biomedical materials research part a
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.849
H-Index - 150
eISSN - 1552-4965
pISSN - 1549-3296
DOI - 10.1002/jbm.a.34357
Subject(s) - lysozyme , protein adsorption , adsorption , materials science , biomaterial , kinetics , biophysics , layer (electronics) , contact angle , chemical engineering , chromatography , nanotechnology , biochemistry , chemistry , composite material , organic chemistry , biology , physics , quantum mechanics , engineering
A new process has been developed to determine the biological activity of an intact layer of lysozyme deposited onto a biomaterial surface. This process is applied to a number of common hydrogel contact lenses. The activity of the surface‐adsorbed protein is measured using a standard micrococcal activity assay, with extra steps to distinguish between protein on the surface and protein in solution. This is in contrast to protein extraction work in which the activity of all adsorbed protein is measured. For ionic materials, which are known to deposit large amounts of protein, particularly positively charged proteins such as lysozyme, there is evidence for loosely bound protein re‐entering the solution, thus making it impossible to truly separate out the surface‐adsorbed protein. This optimized process provides the first quantification of the biological activity of an intact layer of surface‐adsorbed protein at a hydrogel interface. © 2012 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 101A: 755–764, 2013.