Use of surface plasmon resonance biosensor technology as a possible alternative to detect differences in binding of enantiomeric drug compounds to immobilized albumins.
The use of biosensors for monitoring real time interactions between biomolecules and drug compounds has a lot of advantages over presently existing detection methods, the major ones being the elimination of radio labels and rapid screening. We can also obtain information about the kinetic parameters and these values may serve as useful indicators towards subtle differences in the binding strength and characteristics of closely related drug compounds and enantiomers. The Biacore 3000 biosensor based on the Surface Plasmon Resonance (SPR) technology was used to assess the albumin protein binding differences between two enantiomers of a drug compound. Normalized responses (NRU) and affinity constants (K(D)) were readily calculated. Statistical parameters like mean normalized responses, %CV values were determined to make the technique robust. The %CV values obtained were within the preset limits of < or = 25% (FDA limits for drug development and method validation protocols) for the binding interactions for majority of the concentrations studied. For example, the %CV values for the normalized responses for the binding of the control drug warfarin to human albumin ranged from 7.9 to 24.3%. The method gave reproducible results, and the results indicated slight differences in binding patterns of the enantiomers to human and rat albumin.
Ahmad
Ateeq
A
Global Metabolism Investigative Sciences, Pharmacia Corporation, 4901 Searle Parkway, Skokie, IL 60077, USA. aahmad@neophrm.com
Ramakrishnan
Anand
A
McLean
Matthew A
MA
Breau
Alan P
AP
D000818:Animals; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods*; D005240:Feasibility Studies; D006801:Humans; D007536:Isomerism; D046911:Macromolecular Substances; D004364:Pharmaceutical Preparations / Q000032:analysis / Q000737:chemistry* / Q000145:classification*; D011485:Protein Binding; D051381:Rats; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012709:Serum Albumin / Q000737:chemistry*; D020349:Surface Plasmon Resonance / Q000295:instrumentation* / Q000379:methods*
D046911:Macromolecular Substances; D004364:Pharmaceutical Preparations; D012709:Serum Albumin