Glycan profiling utilizing capillary electrophoresis

Abstract

A method for glycan profiling by capillary electrophoresis (CE), and a CE system for glycan analysis (N-Glycan). The CE system uses integrated dual optical fibers for both radiation excitation and emission detection. The CE system is configured for performing a two-color detection for data analysis. A single radiation excitation source is used to excite two emission fluorophores or dyes in the sample solution to be analyzed. One emission dye is to tag the sample and the other dye is used to provide a reference marker (e.g., a Dextran Ladder) for the sample run. Two detectors (e.g., photomultipler tubes (PMTs)) are applied to simultaneously detect the fluorescent emissions from the dyes. The data collected by both detectors are correlated (e.g., synchronized, and / or super-positioned for analysis) for accurate data peak identification.

Description

PRIORITY CLAIM[0001]This application claims the priority of U.S. Provisional Patent Application No. 62 / 002,142 filed on May 22, 2014. This provisional patent and all documents discussed below are fully incorporated by reference as if fully set forth herein.BACKGROUND OF THE INVENTION[0002]1. Field of the Invention[0003]The present invention relates to bio-analysis, in particular a method for glycan profiling by capillary electrophoresis and a capillary electrophoresis instrument for performing the method.[0004]2. Description of Related Art[0005]Post-translational modification of proteins with oligosaccharides to form glycoproteins is a common biological motif. These glycoprotein oligosaccharides are involved in a wide range of biological and physiological processes including recognition and regulatory functions, cellular communication, gene expression, cellular immunity, growth, and development. Aberrant glycosylation of proteins is connected to cancer progression, invasion, and met...

AI Technical Summary

Benefits of technology

The present invention provides a new method and system for analyzing glycans, which are important components of glycoproteins, using a high-speed, cost-effective capillary gel electrophoresis system with integrated dual fiber optic detection technology. This system can hold multiple samples and quickly analyze them, saving time and ensuring accurate results. The system also uses a two-color detection method, which allows for accurate data peak identification and sizing for glycan profiling. Overall, this invention simplifies and shortens sample separation and detection into a single run and provides reproducible peak identification and sizing for glycan profiling.

Problems solved by technology

In view of this heterogeneity and the presence of branched structures, the analysis of glycans is much more complicated than protein chemistry.

Although this technique is sensitive in the sub-pico molar range, the resolution between the glycans can be poor due to the limitation on the size of the gel.

However, slab gel electrophoresis for bio-analysis is labor intensive and needs to be drastically improved in terms of resolving power, throughput and cost per sample.

However, the current selling price for CE-based LIF systems is much more expensive than traditional slab-gel based bio-analysis systems due to the complicated optical detection mechanism.

The expensive CE-based systems are thus out of reach for all but a few well-funded laboratories and seems to be a high-cost barrier.

Further, CE is commonly avoided in routine analysis because it is reputed to be a troublesome technique with high failure rates.

However this is no longer true because instrument manufacturers have drastically improved instrument design and overall CE knowledge has increased.

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