The Thermo Scientific Vanquish UHPLC system with Fluorescence detection is applied for the rapid glyco-profiling of a commercial chimeric IgG1 mAb.The separation is performed on a Thermo Scientific Accucore-150-Amide HILIC column in under 2.5 minutes. The developed methodology offers an excellent analytical platform for ultrafast glycan profile comparison of different batches​ of these mAbs.

The Thermo Scientific Vanquish H system with fluorescence detection is applied for the rapid glycoprofiling of a commercial chimeric IgG1 mAb. The separation is performed on a Thermo Scientific Accucore-150-Amide HILIC column in under 2.5 minutes. Developed methodology offers an excellent analytical platform for ultrafast glycan profile comparison of various production batches.

The Thermo Scientific Vanquish H UHPLC system was used for the high resolution determination of 2-AA (anthranilic acid) labelled N-glycans released from a commercial chimeric IgG1 mAb (Infliximab). The separation is performed on a 150 mm Accucore 150-Amide-HILIC column with fluorescence detection giving separation in less than 30 minutes.N-Glycans present at the asparagine 297 residue in the CH2 domain of the Fc region of IgG play a crucial role in modulating the structural stability and functional activity relationship of the antibody.

The Thermo Scientific Vanquish H UHPLC system is applied for the rapid determination of 2-AB (2-aminobenzamide) labelled N-glycans released from human serum IgG in less than 15 minutes using a 50 mm Thermo Scientific Accucore 150-Amide-HILIC column with fluorescence detection. The method is able to determine N-Glycans present at the asparagine 297 residue in the CH2 domain of the Fc region of IgG that play a crucial role in modulating the structural stability and functional activity relationship of the antibody.

This application update demonstrates improved chromatographic separation achieved using a Thermo Scientific™ Dionex™ CarboPac™ PA200 column for the HPAE-FLD method described in proposed United States Pharmacopeia (USP) General Chapter <212> to separate 2-AB-labeled N-linked oligosaccharides from human α1 acid-glycoprotein. An ICS-6000 can be used for this application.

Here, oligosaccharides released from two glycoproteins are labeled with a fluorophore followed by their purification and HPAE-FLD analysis. Two different methods for purification of the labeled oligosaccharides from unbound labeling reagent were tested along with two elution conditions. The data show that the variations tested here do not significantly affect the retention time profiles and that the method described here is suitable for routine profiling of oligosaccharides released from glycoproteins. An ICS-6000 can be used for this application.

Here, oligosaccharides released from two glycoproteins are labeled with a fluorophore followed by their purification and HPAE-FLD analysis. Two different methods for purification of the labeled oligosaccharides from unbound labeling reagent were tested along with two elution conditions. The data show that the variations tested here do not significantly affect the retention time profiles and that the method described here is suitable for routine profiling of oligosaccharides released from glycoproteins. An ICS-6000 can be used for this application.

HPLC of 2-aminobenzamide (2-AB) labeled glycans with fluorescence detection is one of the major analyses in biotherapeutics characterization. Here we show the separation of 2-AB-glycans originated by bovine fetuin and human IgG with The Thermo Scientific Vanquish Flex UHPLC. The Thermo Scientific GlycanPac AXR-1 column is used for the fetuin glycans. The Thermo Scientific Accucore 150-Amide-HILIC is used for the IgG glycans.

The Thermo Scientific nanoLC system is applied for the analysis of 2-aminobenzamide labeled dextran ladder. The separation is performed on a Thermo Scientific Accucore 150-Amide-HILIC nanoLC column, with UV detection at 240 and 330 nm.

The Thermo Scientific Dionex Ultimate 3000 BioRS system in combination with the Thermo Scientific Accucore 150-Amide-HILIC is used to run separation of 2-AB-labelled dextran ladder in gradient mode. The column compartment was set at different temperature between 20 and 50 °C. At high temperature the selectivity between oligomers was improved, and peaks were in average narrower. Consequently higher peak capacity was achieved at high temperature.

The Thermo Scientific Dionex UltiMate 3000 BioRS is applied for separation of 2-aminobenzamide labeled dextran ladder. The HILIC separation is achieved with Thermo Scientific Accucore 150-Amide-HILIC 2.6 µm, 150 x 2.1 mm.The Glucose Units (GU) values are easily assigned the dextran ladder peaks. A calibration curve logarithm GU versus retention time is automatically created with Thermo Scientific Dionex Chromeleon 7.2 Chromatography Data System. The calibration curve is used to assign GU value to unknown glycans.

The analysis of 2AB labelled bovine fetuin glycans was performed on the Thermo Scientific Accucore 150-Amide-HILIC HPLC column. The analysis was simple and robust, leading to the separation and detection of eight individual glycan peaks.

The Thermo Scientific UltiMate 3000 RSLC system is applied for the analysis of a 2-aminobenzamide labelled dextran ladder. The separation is performed on a Thermo Scientific Accucore 150-Amide-HILIC HPLC column, with fluorescence detection.