glyXalign: High‐throughput migration time alignment preprocessing of electrophoretic data retrieved via multiplexed capillary gel electrophoresis with laser‐induced fluorescence detection‐based glycoprofiling

Abstract

Glycomics has become a rapidly emerging field and monitoring of protein glycosylation is needed to ensure quality and consistency during production processes of biologicals such as therapeutic antibodies or vaccines. Glycoanalysis via multiplexed CGE with LIF detection (xCGE-LIF) represents a powerful technique featuring high resolution, high sensitivity as well as high-throughput performance. However, sample data retrieved from this method exhibit challenges for downstream computational analysis due to intersample migration time shifts as well as stretching and compression of electropherograms. Here, we present glyXalign, a freely available and easy-to-use software package to automatically correct for distortions in xCGE-LIF based glycan data. We demonstrate its ability to outperform conventional algorithms such as dynamic time warping and correlation optimized warping in terms of processing time and alignment accuracy for high-resolution datasets. Built upon a set of rapid algorithms, the tool includes an intuitive graphical user interface and allows full control over all parameters. Additionally, it visualizes the alignment process and enables the user to readjust misaligned results. Software and documentation are available at http://www.glyxera.com.

Comparative Performance of Four Methods for High-throughput Glycosylation Analysis of Immunoglobulin G in Genetic and Epidemiological Research

“…GlyXtool was used for structural identification by patented migration time normalization to an internal standard and N -glycan database driven peak annotation (40). The data comparison was performed by glyXalign (41). …”

High-throughput glycosylation analysis of therapeutic immunoglobulin G by capillary gel electrophoresis using a DNA analyzer

“…[46][47][48] We developed a high-throughput method for the glycosylation analysis of recombinantly expressed IgGs by capillary gel electrophoresis using a DNA analyzer. The highthroughput analysis is achieved by a high degree of automation and by multiplexing that allows up to 96 samples to be analyzed concurrently.…”

Database search assisted N-glycan structure identification

“…A frequent criticism against the GU value based structural elucidation of glycans is that the freely available apps have no ability to process multiple runs at the same time in an automated fashion. While the glyXera suite supports multilane CE migration time alignment for capillary gel electrophoresis-based N-glycan analysis [43] unfortunately, it does not address GU value calculation or direct structural elucidation. For a description of glycan structure and glycoprotein centric databases readers are referred to GlyGen [44] and GlyCosmos [45] activities.…”