Total serum protein N‐glycome profiling on a capillary electrophoresis‐microfluidics platform

Abstract

We implemented 8-aminopyrene-1,3,6-trisulfonic acid (APTS)-labeled asparagine-linked glycan (N-glycan) profiling on a microfluidic electrophoresis platform. Using 11.5 cm effective length etched channels and 4% linear polyacrylamide as the separation matrix, the major N-glycans in human serum were profiled in 12 min with a resolution comparable to what is achieved for these analytes on gel-based DNA sequencers. This demonstration suggests a practical clinical application for high-speed compact analyzers which might be uniquely based on microfluidic devices.

Comparison of methods for the analysis of therapeutic immunoglobulin G Fc-glycosylation profiles—Part 1: Separation-based methods

“…28,[39][40][41][42] All other methods applied fluorescent labeling. Four methods used electrophoretic separation, which included conventional high-resolution capillary gel electrophoresis with laser-induced fluorescence [CE-LIF(APTS-HR1)], [43][44][45][46][47][48] DNA-sequencer-aided fluorophore-assisted carbohydrate electrophoresis after 8-aminopyrene-1,3,6-trisulfonic acid (APTS) labeling for high-throughput screening [DSA-FACE(APTS)], [49][50][51][52][53][54] high-resolution capillary gel electrophoresis with rapid labeling with APTS via reductive amination [CE-LIF (APTS-HR2)], and cartridge-based capillary gel electrophoresis with rapid 8-aminonaphthalene-1,3,6-trisulfonate (ANTS) labeling, in development specifically for screening [CCGE(ANTS)]. The CE methods differ with regard to the labeling method: APTS labeling for the "normal" method requires 4 to 24 h for labeling, whereas rapid reductive amination has been optimized Three laboratories were involved in performing the experiments, an analytical laboratory in a development department, a quality control laboratory and a laboratory of a vendor of glycoanalytical tools.…”

Protein attachment onto silica surfaces – a survey of molecular fundamentals, resulting effects and novel preventive strategies in CE

“…Due to its unique properties, CE becomes increasingly implemented in the ''omics'' field of research [3,4], including, e.g. genomics [5,6], metabolomics [7][8][9][10], glycomics [11][12][13] and particularly proteomics [14,15]. Although peak capacity of CE cannot compete with 2-DE approaches, which easily distinguish 1000-3000 and theoretically up to 10 000 protein spots [16][17][18], CE provides an unrivalled selectivity when separating complex mixtures of closely related proteins, differing only slightly in their primary sequence [19], in their PTMs, encompassing, e.g.…”

Chip‐based CE for rapid separation of 8‐aminopyrene‐1,3,6‐trisulfonic acid (APTS) derivatized glycans

“…In comparison to separations obtained in standard CE and/or the previously reported high-resolution separation on a chip with long (22 cm) separation channel [19] it is clear that the resolution is lower. This might be a handicap for some future applications.…”