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SOLAµ for Pre- Analysis Sample Concentration
Jon Bardsley and, Ken Meadows, Thermo Fisher Scientific
This application note demonstrates the use of Thermo Scientific™ SOLA�™ SPE product to enhance sample pre-concentration prior to analysis. Additional benefits include reduced workflow and stability for analytes susceptible to loss or degradation during evaporation and reconstitution. The use of a Thermo Scientific™ Accucore™ HPLC column provided fast and efficient separation without the need for an ultra high pressure system. MS-/MS detection was performed on a Thermo Scientific™ TSQ Vantage™ mass spectrometer. Read More |
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SOLAµ for Reduced Sample Volume Analysis
Jon Bardsley and Ken Meadows, Thermo Fisher Scientific
This application note demonstrates how the Thermo Scientific™ SOLAµ™ SPE product can be used to facilitate the scale down of an extraction method for use when sample volume is limited. The use of a Thermo Scientific™ Accucore™ HPLC column provided fast and efficient separation without the need for an ultra high pressure system. MS-MS detection was performed on a Thermo Scientific™ TSQ Vantage™ mass spectrometer. Read More |
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Improving Sensitivity with SOLAμ for Analytes Susceptible to Issues During Pre-Concentration Dry Down
Jon Bardsley and Ken Meadows, Thermo Fisher Scientific
This application note demonstrates the use of Thermo Scientific™ SOLAµ™ SPE product for the extraction analytes which are susceptible to loss or degradation during evaporation and reconstitution. The use of a Thermo Scientific™ Accucore™ HPLC column provided fast and efficient separation without the need for an ultra high pressure system. MS-/MS detection was performed on a Thermo Scientific™ TSQ Vantage™ mass spectrometer. Read More |
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Separation of 2AB-Labeled N-Linked Glycans from Bovine Fetuin on a Novel Ultra High Resolution Mixed-Mode Column
Udayanath Aich, Julian Saba, Xiaodong Liu, Jim Thayer, and Chris Pohl, Thermo Fisher Scientific
This application note demonstrates the separation of 2AB-labeled N-linked glycans released from proteins (bovine fetuin) by reversed phase (RP) / weak anion exchange (WAX) mixed-mode chromatography. The separation is carried out with both 1.9 �m and 3 µm Thermo Scientific™ GlycanPac™ AXR-1 columns using fluorescence detection. The method exhibits an excellent separation based on charge, isomeric structure, and size. Read More |
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Separation of 2AA-Labeled N-Linked Glycans from Glycoproteins on a High Resolution Mixed-Mode Column
Udayanath Aich, Julian Saba, Xiaodong Liu, Jeff Rohrer, Jim Thayer, and Chris Pohl, Thermo Fisher Scientific
This application note demonstrates the separation of 2AA-labeled N-linked glycans released from bovine fetuin using a reversed-phase / weak anion-exchange mixed-mode column (Thermo Scientific™ GlycanPac™ AXR-1, 1.9 µm, 150 � 2.1 mm) with fluorescence detection. The method exhibits excellent separation of glycans based on charge, isomeric structure, and size. Read More |
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Structural Analysis of Native N-Glycans Released from Proteins Using a Novel Mixed-Mode Column and a Hybrid Quadrupole-Orbitrap Mass Spectrometer
Udayanath Aich, Julian Saba, Xiaodong Liu, and Chris Pohl, Thermo Fisher Scientific
This application note compares two approaches for the structural characterization of N-glycans released from proteins. Analysis was carried out using a novel Thermo Scientific™ GlycanPac™ AXH-1 column and a Thermo Scientific™ Q Exactive™ hybrid quadrupole-Orbitrap™ mass spectrometer. Read More |
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A Fast and Robust Linear pH Gradient Separation Platform for Monoclonal Antibody (mAb) Charge Variant Analysis
Shanhua Lin, Julia Baek, and Chris Pohl, Thermo Fisher Scientific
This application note describes a fast, 10 min cycle time separation of mAb charge variants using a linear pH gradient separation method. A gradient from pH 5.6 to pH 10.2 was generated over time by running a linear pump gradient from 100% Thermo Scientific™ CX-1 pH gradient buffer A (pH 5.6) to 100% CX-1 pH gradient buffer B (pH 10.2). Linear UV response with up to 300 μg of protein loading was achieved. The elution pH values of mAbs are in linear relationship with their corresponding pI values. Read More |
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Analysis of Monoclonal Antibodies and Their Fragments by Size-Exclusion Chromatography Coupled with an Orbitrap Mass Spectrometer
Shanhua Lin, Hongxia Wang, Zhiqi Hao, and Xiaodong Liu, Thermo Fisher Scientific
The Thermo Scientific™ MAbPac™ SEC-1 column uses size-exclusion chromatography (SEC) and is designed for monoclonal antibody (mAb) analysis, including monomers, aggregates, and fragments. Coupling size-exclusion chromatography with a high-resolution Thermo Scientific™ Exactive™ Plus EMR Orbitrap™ mass spectrometer (SEC-MS) enables accurate mass measurement of mAb and its fragments in either native or denatured state. When using a non-denaturing, volatile eluent, such as 20 mM ammonium formate, the intact mass of mAb can be measured in its native state. When using denaturing volatile eluents, such as 20% acetonitrile, 0.1% formic acid, and 0.05% trifluoroacetic acid, mAb heavy chain (HC), light chain (LC), Fab, and Fc fragments are successfully separated on MAbPac SEC-1 and their masses are determined accurately. Read More |
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