An application of ultraviolet spectroscopy to study interactions in proteins solutions at high concentrations

Journal of Pharmaceutical Sciences
Santosh V ThakkarC Russell Middaugh

Abstract

Studies of protein-protein interactions (PPIs), especially in high-concentration solutions, have become increasingly important from a pharmaceutical perspective. Analytical methods used to study protein interactions, however, rely primarily on the detection of nonideality in relatively dilute (<50 mg/mL) solutions. We present here an application of variable-pathlength ultraviolet (UV)-visible absorption spectroscopy to examine and better understand such interactions over a wide concentration range (5-240 mg/mL) using several representative proteins. In this study, the change in UV absorption (or extinction coefficient) was monitored by determining delta absorbance (ΔAbs), the difference between the measured absorbance and the corresponding theoretical absorbance (calculated from gravimetric dilution), over a wide range of protein concentrations. The ΔAbs, corrected for light scattering, was found to increase with protein concentration for three model proteins (bovine serum albumin, lysozyme, and monoclonal antibody). Because PPIs influence solution viscosity, we studied the correlation between ΔAbs measurements and viscosity as a function of protein concentration. The magnitude of ΔAbs and solution viscosity followed similar tr...Continue Reading

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Citations

Dec 15, 2015·Organic & Biomolecular Chemistry·Kapil UpadhyayaRama Pati Tripathi
Nov 15, 2016·Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology·Otávio A ChavesCarlos Serpa
Sep 22, 2018·Biotechnology Progress·William S McKechnieSunitha Kandula
Nov 8, 2018·BioMed Research International·Olga IlinskayaYuriy Zuev
Jul 21, 2020·Biotechnology and Bioengineering·Melissa HolsteinZheng Jian Li

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