Final answer:
The question pertains to laboratory methods used to measure protein binding percentages, particularly the binding affinity between albumin and the antibiotic levofloxacin. Methods include high throughput screening, equilibrium dialysis, ultrafiltration, HPLC, and LC-MS/MS, with fluorescence used for sensitive optical measurement.
Step-by-step explanation:
Laboratories utilize several methods to measure protein binding affinity, particularly the binding of drugs to plasma proteins like albumin. Often, labs will use high throughput screening (HTS) ultrafiltration methods with a 96-well plate to evaluate the binding of plasma proteins. One specific example is measuring the binding affinity between albumin and levofloxacin, a common antibiotic. Protein binding assessments are critical in understanding the bioavailability and distribution of drugs within the body. Equilibrium dialysis and ultrafiltration are frequently used during initial research phases to determine the percentage of a drug that remains unbound to plasma proteins.
For quantitative analysis, techniques such as high-performance liquid chromatography (HPLC) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) are often employed. These techniques determine the concentration of unbound drug prior to and after ultrafiltration, providing data on the extent of protein binding. The sensitive optical measurement of fluorescence is also used to study the details of antibiotic binding to albumin.