Fluidics-resolved estimation of protein adsorption kinetics in a biomicrofluidic system.
Protein adsorption on surfaces is a complex phenomenon that is described by the balance of convective/diffusive transport of the protein species to the surface and its adsorption/desorption at the surface. The extent of binding depends on a variety of factors such as protein/surface interactions, availability of binding sites, localized concentrations of protein near biomaterial surfaces and flow characteristics of the protein in that region. Factors such as time-varying flows, complex device geometries, presence of multiple competitive species, or possible denaturing of proteins when they attach to the surface make it extremely difficult to quantitatively analyze protein interactions with surfaces. Adsorption/desorption rate constants are often inferred using simplistic models which neglect mass transport and have limited use across different microfluidic systems and flow protocols. In this work, we have developed and demonstrated a fluidics-resolved model that evaluates protein adsorption, accounting for both the fluidic transport and the biochemical kinetics in complex biomicrofluidic devices. The model is valid for both flow and static conditions. An automated procedure was also developed to extract the "intrinsic" mass-transport-independent adsorption kinetic rate constants from experimental data using a least squares optimization method. The automated data extraction methodology is applied to two proteins (alkaline phosphatase and glucose oxidase) that have been brought into contact with poly(etheretherketone) and Teflon capillaries. The applicability of the procedure in analyzing flow and adsorption in complex microfluidic structures is also demonstrated.
Jenkins
J
J
CFD Research Corp. Huntsville, AL 35805, USA.
Prabhakarpandian
B
B
Lenghaus
K
K
Hickman
J
J
Sundaram
S
S
D000327:Adsorption; D000469:Alkaline Phosphatase / Q000737:chemistry*; D005949:Glucose Oxidase / Q000737:chemistry; D007700:Kinetics; D044085:Microfluidics
D005949:Glucose Oxidase; D000469:Alkaline Phosphatase