Abstract

FR.21.01

Protein biomarker as a new diagnostic approach in ophthalmology

Grus F. H. 
Experimental Ophthalmology, Department of Ophthalomology, University of Mainz

Objective: Biomarker studies and proteomics analyses play an important role for early detection of diseases, but also for the development of new treatment options. Proteomic profiling studies e.g. by ProteinChips (Seldi-TOF) can improve the understanding of the pathogenesis of disease processes and can lead to earlier diagnosis. In the area of cancer diseases, first diagnostic approaches are nearly ready for clinical routine (e.g. ovarial cancer). In this paper, the chances of biomarker profiling in Ophthalmology will be presented and discussed. Especially the analysis of eye fluids such as tear film needs the use of techniques which are sensitive enough to detect even low abundant proteins in the small tear volumes. Therefore, we use high-sensitive mass spectrometry based techniques. Different biomarker profiling studies in glaucoma and ocular surface diseases will be presented.
Methods: Antibody- and protein profiling studies by means of Protein-Chip analysis were performed in glaucoma and ocular surface diseases. Sera and tears were fractionated on an anion exchange resin using stepwise pH gradients, and subsequently analyzed on three different types of ProteinChip arrays. Proteins which bound to the surface were detected by surface enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS). Peak mass and normalized intensity were used for statistical analysis.
Results: Complex protein patterns were found in all patients. More than 4000 protein clusters were identified across the three different chip surfaces, fractions, and laser energy settings. Multivariate analysis of discriminance can test for statistical differences between the subgroups using the entire complex staining pattern for the calculation. The method successfully found a significant difference between all subgroups. Based on 8 biomarker panel including the most significant biomarkers between all groups, these panels were tested for sensitivity and specificity.
Conclusions: Biomarker analysis might be a very promising approach for early-detection and could lead to novel drug targets in future.

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