AVS 45th International Symposium
    Biomaterial Interfaces Group Monday Sessions
       Session BI-MoM

Paper BI-MoM9
Qualitative and Quantitative Mass Spectrometric Methods for Probing Surface-Protein Binding Affinity

Monday, November 2, 1998, 11:00 am, Room 326

Session: Protein Solid-Surface Interactions
Presenter: G.R. Kinsel, University of Texas, Arlington
Authors: G.R. Kinsel, University of Texas, Arlington
A.K. Walker, University of Texas, Arlington
L. Chen, University of Texas, Arlington
K.D. Nelson, University of Texas, Arlington
Y. Wu, University of Texas, Arlington
R.B. Timmons, University of Texas, Arlington
Correspondent: Click to Email
We have recently shown that Matrix Assisted Laser Desorption Ionization (MALDI) Mass Spectrometry (MS) can be used as an efficient tool for characterizing surface protein interactions. When using a standard sample preparation methodology there is a direct inverse correlation between the surface-protein binding affinity and the magnitude of the protein MALDI MS ion signal. The MALDI MS method offers numerous advantages over conventional bioanalytical methods for assaying surface-protein binding including: 1) surface binding of a broad spectrum of proteins and biomolecules can be assayed, 2) no modification (e.g. radiolabeling, fluorescent labeling) of the protein is required, 3) analysis is straightforward and can be completed in under 30 minutes 4) sensitivities are comparable to radiolabeled protein binding studies. Our present research is focused on assessment of the conditions under which the MALDI ion signal can be quantitatively correlated to the surface-protein binding affinity. Specifically we have examined the impact of changes in a variety of surface (morphology, chemical functionality, etc) and protein solution (protein choice, solution pH, etc.) characteristics on the general quantitative MALDI ion signal / binding affinity correlation. These studies reveal both the range of conditions across which the MALDI MS methodology can be applied to quantitate surface-protein binding as well as insights into the relative impact of various surface / solution parameters on the adsorption process.