Evaluating SPR array imaging for glycobiology

Lead Research Organisation: John Innes Centre
Department Name: Contracts Office

Abstract

Whilst carbohydrate structure and recognition are clearly central to biology, our appreciation of their structure and function is still rudimentary compared to proteins and nucleic acids. It has been estimated that around 2 percent of the human genome is dedicated to glyco-active enzymes (up to 6 percent in plants) and that over 70 percent of all proteins in man are glycosylated. In turn this has led to a need for analysis of larger numbers of glycan structures, and their cognate binding proteins, which has provided the impetus for the development of new technologies with high-throughput potential. Latterly, glycoscientists have identified carbohydrate microarrays (glycoarrays) as a key tool for the high-throughput studies that are necessary to understand this complex area of biology. The purpose of this study is to evaluate new methods with which to analyse carbohydrate-active enzymes based on integrated, chip-based surface plasmon resonance spectroscopy-mass spectrometry. This pilot project will investigate the potential of SPR imaging (SPRi)-MALDI-tof mass spectrometry for the analysis of chip-based glycan synthesis. The aims of this project are two-fold: A) To integrate MALDI-tof and SPRi analysis on the same carbohydrate microarray; and B) To assess the suitability of a commercial SPRi instrument and sensor chips for this purpose. This study will involve: 1. Assess protein-carbohydrate interactions with SPR imaging of a glycan microarray and plant lectins in solution. 2. Assess step-wise enzymatic synthesis of glycans on-chip by MALDI-tof and SPRi. 3. Assess enzymatic polymerisation on-chip by MALDI-tof and SPRi. 4. Assess enzymatic hydrolysis on-chip by MALDI-tof and SPRi.

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