Analysis of using I125 radiolabeling for quantifying protein on contact lenses
Hall, B, Heynen, H, Jones, L, Forrest, J. Analysis of using I125 radiolabeling for quantifying protein on contact lenses. Current Eye Research 2016;41(4):456-465.
To investigate the accuracy of I125 radiolabeling to quantitatively determine the deposition of protein onto various commercially available contact lens (CL) materials.
Commercially available silicone hydrogel and conventional hydrogel CL materials were examined for times ranging from 10 s to 1 week. Adsorption of free I125 was measured directly for the CL. The use of dialyzing labeled proteins and/or using NaI to compete with free I125 uptake was investigated as ways to minimize effects due to free I125.
At all time points and with all lens materials, there was 0.3 μg/lens or greater of apparent mass attributable to free I125 uptake. Dialyzing labeled proteins significantly reduced free I125 uptake for all materials investigated. The benefit of using dialyzed protein was most prominent at shorter times, as free I125 is continuously generated over time. Using NaI can reduce free I125 uptake for some lens materials, but this is shown to directly affect protein deposition on some materials.
Periodic replenishment of incubation solutions with freshly dialyzed labeled protein to limit free I125 generation is recommended, but the incorporation of NaI onto the buffer solution is not. Irrespective of the exact procedure to limit free I125 uptake, extra steps must be performed to quantify the amount of I125 adsorbed onto contact lens materials, to determine thresholds of confidence with respect to the actual protein deposition that occurs.