Recognition of conformational changes in β-lactoglobulin by molecularly-imprinted thin films

AGRO 116

David W. Britt, dbritt@cc.usu.edu1, Nicholas Turner2, Xiao Liu1, Sergey Piletsky2, and Vladimir Hlady, vladimir.hlady@m.cc.utah.edu3. (1) Dept. of Biological Engineering, Utah State University, 4105 Old Mail Hill, Logan, UT 84322-4105, (2) Cranfield Health, Cranfield University, Bedfordshire, Silsoe, MK45 4DT, United Kingdom, (3) Department of Bioengineering, University of Utah, 50 S. Central Campus Dr., Rm. 2480, Salt Lake City, UT 84112
Pathogenesis in protein conformational diseases is initiated by changes in protein secondary structure. This molecular restructuring presents an opportunity for novel shape-based detection approaches, as protein molecular weight and chemistry are otherwise unaltered. Here we apply molecular imprinting to discriminate between distinct conformations of the model protein β-lactoglobulin (BLG). Thermal and fluoro-alcohol induced BLG isoforms were imprinted in thin films of 3-aminophenylboronic acid on quartz crystal microbalance chips. Enhanced rebinding of the template isoform was observed in all cases when compared to the binding of non-template isoforms over the concentration range 1-100 µg/ml. Furthermore, it was observed that the greater the changes in the secondary structure of the template protein the lower the binding of native BLG challenges to the imprint, suggesting a strong steric influence in the recognition system. This feasibility study is a first demonstration of molecular imprints for recognition of distinct conformations of the same protein.
 

Nanotechnology in Agriculture
8:30 AM-11:40 AM, Wednesday, August 22, 2007 BCEC -- 259A, Oral

Division of Agrochemicals

The 234th ACS National Meeting, Boston, MA, August 19-23, 2007