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ABSTRACT: Purpose
To measure the hydrodynamic radii of intravitreal anti-VEGF drugs ranibizumab, aflibercept and bevacizumab with ?s time-resolved phosphorescence anisotropy.Methods
Ruthenium-based dye Ru(bpy)2(mcbpy?-?O?-?Su?-?ester)(PF6)2, whose lifetime of several hundred nanoseconds is comparable to the rotational correlation time of these drugs in buffer, was used as a label. The hydrodynamic radii were calculated from the rotational correlation times of the Ru(bpy)2(mcbpy?-?O?-?Su?-?ester)(PF6)2-labelled drugs obtained with time-resolved phosphorescence anisotropy measurements in buffer/glycerol solutions of varying viscosity.Results
The measured radii of 2.76±0.04 nm for ranibizumab, 3.70±0.03 nm for aflibercept and 4.58±0.01 nm for bevacizumab agree with calculations based on molecular weight and other experimental measurements.Conclusions
Time-resolved phosphorescence anisotropy is a relatively simple and straightforward method that allows experimental measurement of the hydrodynamic radius of individual proteins, and is superior to theoretical calculations which cannot give the required accuracy for a particular protein.
SUBMITTER: Hirvonen LM
PROVIDER: S-EPMC4942501 | biostudies-literature | 2016 Aug
REPOSITORIES: biostudies-literature
Hirvonen Liisa M LM Fruhwirth Gilbert O GO Srikantha Nishanthan N Barber Matthew J MJ Neffendorf James E JE Suhling Klaus K Jackson Timothy L TL
Pharmaceutical research 20160525 8
<h4>Purpose</h4>To measure the hydrodynamic radii of intravitreal anti-VEGF drugs ranibizumab, aflibercept and bevacizumab with μs time-resolved phosphorescence anisotropy.<h4>Methods</h4>Ruthenium-based dye Ru(bpy)2(mcbpy - O - Su - ester)(PF6)2, whose lifetime of several hundred nanoseconds is comparable to the rotational correlation time of these drugs in buffer, was used as a label. The hydrodynamic radii were calculated from the rotational correlation times of the Ru(bpy)2(mcbpy - O - Su - ...[more]