Multi-Site N-glycan mapping study 1: Capillary electrophoresis – laser induced fluorescence
نویسندگان
چکیده
An international team that included 20 independent laboratories from biopharmaceutical companies, universities, analytical contract laboratories and national authorities in the United States, Europe and Asia was formed to evaluate the reproducibility of sample preparation and analysis of N-glycans using capillary electrophoresis of 8-aminopyrene-1,3,6-trisulfonic acid (APTS)-labeled glycans with laser induced fluorescence (CE-LIF) detection (16 sites) and ultra high-performance liquid chromatography (UHPLC, 12 sites; results to be reported in a subsequent publication). All participants used the same lot of chemicals, samples, reagents, and columns/capillaries to run their assays. Migration time, peak area and peak area percent values were determined for all peaks with >0.1% peak area. Our results demonstrated low variability and high reproducibility, both, within any given site as well across all sites, which indicates that a standard N-glycan analysis platform appropriate for general use (clone selection, process development, lot release, etc.) within the industry can be established.
منابع مشابه
Restriction mapping of genes by capillary electrophoresis with laser-induced fluorescence detection.
Restriction mapping is one of the essential steps in gene analysis and molecular biology studies. Slab gel electrophoresis is the traditional way to separate DNA fragments for restriction mapping. However, slab gel electrophoresis does not provide sufficient resolution as required in many mapping applications, and the use of radioisotopes in traditional mapping methods creates health hazards. I...
متن کاملUltrasensitive detection and quantification of acidic disaccharides using capillary electrophoresis and quantum dot-based fluorescence resonance energy transfer.
Rapid and highly sensitive detection of the carbohydrate components of glycoconjugates is critical for advancing glycobiology. Fluorescence (or Förster) resonance energy transfer (FRET) is commonly used in detection of DNA, in protein structural biology, and in protease assays but is less frequently applied to glycan analysis due to difficulties in inserting two fluorescent tags into small glyc...
متن کاملHigh-sensitivity laser-induced fluorescence detection for capillary electrophoresis.
Capillary electrophoresis provides very high efficiency separations of biological molecules. Laser-induced fluorescence produces very high sensitivity detection. The combination of the two techniques results in an analytical tool with unprecedented properties: separations with more than a million theoretical plates and detection limits of a few hundred analyte molecules. This paper considers th...
متن کاملCapillary electrophoresis with wavelength-resolved laser-induced fluorescence detection.
Capillary electrophoresis (CE) enables rapid separations with high separation efficiency and compatibility with small sample volumes. Laser-induced fluorescence detection can result in extremely low limits of detection in CE. Single-channel fluorescence detection, however, furnishes little qualitative information about a species being detected, except for its CE migration time. Use of multidime...
متن کاملMulti-color DNA Analysis in an Optofluidic Chip
Sorting and sizing of DNA molecules by capillary electrophoresis (CE) within the human genome project have enabled the genetic mapping of various illnesses. By the use of miniaturized lab-on-achip devices, on-chip-integrated DNA sequencing and genetic diagnostics have become feasible. The inherent advantages of microfluidic CE separation of differently sized DNA molecules, high-speed operation ...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
دوره 8 شماره
صفحات -
تاریخ انتشار 2016