Glycan profiling utilizing capillary electrophoresis

A method of glycan profiling, comprising:

• providing a separation channel having a first longitudinal axis along which a N-glycan sample undergoes separation into sample components, and a detection zone defined along the separation channel through which the sample components pass;
• prior to subjecting the N-glycan sample to separation, the N-glycan sample is provided with a sample marker corresponding to a first fluorescence emission having a first characteristic wavelength, wherein the sample components are labeled by the sample marker for identification as the N-glycan sample undergoes separation, wherein the N-glycan sample is further provided with a reference marker corresponding to a second fluorescence emission having a second characteristic wavelength different from the first characteristic wavelength, wherein the reference marker provides a reference to facilitate identification of the sample components as the N-glycan sample undergoes separation;
• providing a radiation source that provides an incident radiation to induce the first fluorescence emission from the sample marker having the first characteristic wavelength and the second fluorescence emission from the reference marker having the second characteristic wavelength, and wherein the sample marker comprises a first material that emits the first fluorescence emission when induced by the incident radiation, and the reference marker comprises a second material that emits the second fluorescence emission when induced by the incident radiation;
• providing an incident light guide having a second longitudinal axis, directing the incident radiation from the radiation source to the detection zone along the separation channel;
• providing a first detector detecting the first fluorescence emission having the first characteristic wavelength, and a second detector detecting the second fluorescence emission having the second characteristic wavelength;
• providing an emission light guide having a third longitudinal axis, collecting and directing emitted radiation from the detection zone to a first emission fiber and a second emission fiber, wherein the first emission fiber routes emitted radiation of the first fluorescence emission having the first characteristic wavelength from the emission light guide to the first detector and the second emission fiber routes emitted radiation of the second fluorescence emission having the second characteristic wavelength from the emission light guide to the second detector;
• and
• subjecting the N-glycan sample to high voltage to effect electrophoresis to separate the N-glycan sample into the sample components along the separation channel, wherein the first detector detects the first fluorescence emission and the second detector detects the second fluorescence emission at the detection zone.