Analysis of biomolecular interactions using affinity microcolumns: a review.

Analysis of Free Solute Fractions and Solute-Protein Interactions Using Ultrafast Affinity Extraction and Affinity Microcolumns

Evaluation of silica monoliths in affinity microcolumns for high-throughput analysis of drug-protein interactions.

Silica monoliths in affinity microcolumns were tested for the high-throughput analysis of drug-protein interactions. HSA was used as a model protein for this work, while carbamazepine and R-warfarin were used as model analytes. A comparison of HSA silica monoliths of various lengths indicated columns as short as 1 to 3 mm could be used to provide reproducible estimates of retention factors or p...

Development of affinity microcolumns for drug-protein binding studies in personalized medicine: interactions of sulfonylurea drugs with in vivo glycated human serum albumin.

This report used high-performance affinity microcolumns to examine the changes in binding by sulfonylurea drugs to in vivo glycated HSA that had been isolated from individual patients with diabetes. An immunoextraction approach was developed to isolate HSA and glycated HSA from clinical samples, using only 20 μL of plasma or serum and 6-12 nmol of protein to prepare each affinity microcolumn. I...

Evaluation of affinity microcolumns containing human serum albumin for rapid analysis of drug-protein binding.

This study examined the use of affinity microcolumns as tools for the rapid analysis and high-throughput screening of drug-protein binding. The protein used was immobilized human serum albumin (HSA) and the model analytes were warfarin and L-tryptophan, two solutes often used as site-specific probes for drug binding to Sudlow sites I and II of HSA, respectively. The use of HSA microcolumns in b...

Analysis of drug-protein binding by ultrafast affinity chromatography using immobilized human serum albumin.

Human serum albumin (HSA) was explored for use as a stationary phase and ligand in affinity microcolumns for the ultrafast extraction of free drug fractions and the use of this information for the analysis of drug-protein binding. Warfarin, imipramine, and ibuprofen were used as model analytes in this study. It was found that greater than 95% extraction of all these drugs could be achieved in a...