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Abstract Purpose : Photoreceptor cells are the primary source of vision. Maintaining viable photoreceptor cells is essential for vision. When or before photoreceptors begin to die, even modest enhancement of protein expression at their site may have significant impact on disease progression. The aim of this study is to gain an understanding of naturally occurring expression levels of immuno-proteins like the interphotoreceptor retinoid binding proteins (IRBP202-121 and IRBP204-249) and of the receptors to which they bind in normal control C57BL/6 adult mouse photoreceptor cells. Methods : SPRIGHT-640 B cells (expressing IRBP bound to BSA) used were calibrated to generate calibration curves with a resonance unit range of 1.3-1.9 RU and a concentration range of 0.68-131.07nM over the course of two days. SPRIGHT-648 cells (expressing retinoid receptors) were then used to measure the receptor-ligand binding on day three. The concentrations of bound and unbound substances were determined by subtracting the Respiratory Units (RUs) of the SPRIGHT-648 cells from the SPRIGHT-640 cells. The concentrations of bound IRBP to the receptors were collected, analyzed and calibrated using MATLAB software. Results : Concentrations of bound IRBP to the receptors were determined from the calibration curves obtained from the SPRIGHT-640 and 648 cells. SPRIGHT-640 cells showed a linear relationship between concentrations of IRBP-BSA from 0.68- 132.07 nM and RUs of 1.3-1.9. SPRIGHT-648 cells showed a linear relationship between concentrations of IRBP bound to receptors and RUs of 1.45-1.9. The sedimentation values of the SPRIGHT-640 B cells confirm a linear relationship between sedimentation and concentration of IRBP-BSA. Conclusions : SPRIGHT assays were run to understand the binding affinity between IRBP and its receptor. SPRIGHT-640 calibrations showed the binding affinity of IRBP to the BSA and SPRIGHT-648 calibrations showed the binding affinity between IRBP and its receptors. We were able to confirm a linear relationship between the concentrations of IRBP-BSA and RUs and the concentrations of IRBP bound to its receptors and RUs on SPRIGHT-640 and 648 respectively. We also established a sedimentation change with respect to concentration of IRBP-BSA in SPRIGHT-640 calibrations. This is a 2020 ARVO Annual Meeting abstract.

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