A comprehensive study of concepts and phenomena of the nonspecific adsorption of beta-lactoglobulin.
We investigate nonspecific protein adsorption processes by comparing experimentally measured adsorption kinetics of beta-lactoglobulin with mathematical models. The adsorption and desorption behavior of this protein on a hydrophilic glass surface in citrate buffer (pH 3.0), monitored for a large set of different bulk concentrations (0.5x10(-8) M-1.5x10(-6) M) using a supercritical angle fluorescence (SAF) biosensor, is reported. Increasing adsorption rates and overshootings in the beginning of the adsorption are observed as well as a transition to an almost irreversibly bound state of the protein in the long term. Furthermore, rinsing experiments prove that adsorbed proteins abruptly change their desorption behavior from irreversible to reversible when a critical surface coverage theta(crit) is reached. Based on all experimental observations, a mathematical model composed of three adsorbed states differing in their surface affinity is proposed. Terms to account for lateral interactions between surface-bound proteins are included, which yield an excellent fit of the measured kinetics. For the first time, several phenomena that have been discussed in theoretical studies are confirmed by comparing experimental data with a single model.
Rabe
Michael
M
Institute of Physical Chemistry, University of Zurich, Winterthurerstrasse 190, 8057 Zurich, Switzerland.
Verdes
Dorinel
D
Rankl
Michael
M
Artus
Georg R J
GR
Seeger
Stefan
S
D000327:Adsorption; D015374:Biosensing Techniques / Q000379:methods*; D002021:Buffers; D005456:Fluorescent Dyes / Q000737:chemistry; D005898:Glass / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D007782:Lactoglobulins / Q000737:chemistry*; D008956:Models, Chemical; D013499:Surface Properties
D002021:Buffers; D005456:Fluorescent Dyes; D007782:Lactoglobulins