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Displaying 1-50 of 217 results

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QAR Method for the CarboPac PA300 Column

Instrument Type: IC

This record describes the QAR analysis for a Thermo Scientific Dionex CarboPac PA300 column and provides an eWorkflow for running the analysis..

Charge variant analysis of Ranibizumab sample using volatile buffer and strong cation exchange column.

Instrument Type: HPLC (Biocompatible)

The Thermo Scientific Dionex UltiMate 3000 LC Bio-LC system is applied for the Ion Exchange analysis of ranibizumab sample. The separation was performed on a Thermo Scientific MAbPac SCX-10 with a UV detection at 280 nm.

Fast and Reproducible peptide mapping of an Pertuzumab (IgG1) by using a SMART digest Kit and fully porous 120 A C18 column.

Instrument Type: UHPLC

The Thermo Scientific Dionex Ultimate 3000 system is applied for the peptide mapping analysis of an Pertuzumab (IgG1). The separation was performed on a Thermo Scientific Acclaim RSLC 120 C18 column by using a Thermo Scientific SMART digest kit at UV detection of 214 nm. The SMART Digest kit provides significant improvements in reproducibility, which results in fewer sample failures, higher throughput and the ability to more easily interrogate data..

AN73455: Determination of Nitrite Impurity in Dalteparin Sodium using Ion Chromatography

Instrument Type: IC

In this application note, the nitrite determination described in the USP monograph was evaluated with a Thermo Scientific Dionex IonPac AS15 column using an IC system. The method and conditions were exactly as described in the USP Dalteparin Sodium monograph. Key performance parameters were evaluated including the system suitability separation, linearity, limits of detection, accuracy, and robustness. One Dalteparin Sodium sample was analyzed. The percentage of nitrite result was compared with the USP acceptance criterium.

Ion exchange analysis of teriperatide using polymeric non-porous strong cation exchange column.

Instrument Type: HPLC (Biocompatible)

The Thermo Scientific Dionex UltiMate 3000 LC Bio-LC system is applied for the Ion Exchange analysis of teriperatide. The separation was performed on a Thermo Scientific MAbPac SCX-10 and fluorescence detector.

Chromatographic method for aggregate and fragment analysis of Pertuzumab i.e IgG1 Mab using SEC column.

Instrument Type: HPLC (Biocompatible)

Analysis of protein aggregation and fragment of pertuzumab by size-exclusion chromatography, showing the universal applicability of the Thermo Scientific MAbPac SEC-1 column for aggregate and fragment analysis. Aggregate analysis is major CQA for protein based drugs and requires tight monitoring and control.

Automated biopharmaceutical protein digestion optimization for peptide mapping method development

Instrument Type: Automated Sample Prep

Combining the Thermo Scientific™ SMART Digest™ magnetic kits with the Thermo Scientific™ KingFisher™ Duo Prime purification system provides an automated approach to protein digest method optimization. A robust and reproducible, automated digestion time-course protocol to determine the optimal digest time for a biotherapeutic during development of a peptide mapping method.

Fast and Reproducible peptide mapping of an peptibody (romiplostim) by using a SMART digest Kit and solid core C18 column

Instrument Type: UHPLC

The Thermo Scientific Dionex Ultimate 3000 system is applied for the peptide mapping analysis of an peptibody (Romiplostim). The separation was performed on a Thermo Scientific Vanquish C18+ column by using a Thermo Scientific SMART digest kit at UV detection of 210 nm. The SMART Digest kit provides significant improvements in reproducibility, which results in fewer sample failures, higher throughput and the ability to more easily interrogate data.

TN73348: Carbohydrate Determinations by HPAE-PAD using a PdH Reference Electrode

Instrument Type: IC

We recently introduced a PdH RE for HPAE-PAD carbohydrate determinations. In this work, we demonstrate two popular carbohydrate applications (honey sugars and glycoprotein monosaccharides) using a PdH RE and compare the results to using an Ag/AgCl RE. Our preliminary data indicate that the PdH RE provides comparable results to the standard Ag/AgCl RE for these two applications that use hydroxide eluents.

Chromatographic method for aggregate analysis of Aflibercept i.e fusion protein using SEC column.

Instrument Type: HPLC (Biocompatible)

Analysis of protein aggregation of Aflibercept i.e fusion protein by size-exclusion chromatography, showing the universal applicability of the Thermo Scientific MAbPac SEC-1 column for aggregate analysis of fusion protein. Aggregate analysis is major CQA for protein based drugs and requires tight monitoring and control.

Fast and Reproducible peptide mapping of an Fusion Protein (Aflibercept) by using a SMART digest Kit and solid core C18 column.

Instrument Type: HPLC (Biocompatible)

The Thermo Scientific Dionex Ultimate 3000 system is applied for the peptide mapping analysis of an Fusion Protein (Aflibercept). The separation was performed on a Thermo Scientific Accucore vanquish C18+ column by using a Thermo Scientific SMART digest kit at UV detection of 210 nm. The SMART Digest kit provides significant improvements in reproducibility, which results in fewer sample failures, higher throughput and the ability to more easily interrogate data.

Separation of charge variants of romiplostim by Ion Exchange chromatography using strong cation exchange column

Instrument Type: HPLC (Biocompatible)

The Thermo Scientific Dionex UltiMate 3000 LC Bio-LC system is applied for the Ion Exchange analysis of romiplostim i.e peptibody. The separation was performed on a Thermo Scientific MAbPac SCX-10 with a UV detection at 210 nm.

Oxidised impurity analysis of Denosumab using HIC technique.

Instrument Type: HPLC (Biocompatible)

The Thermo Scientific Dionex ultimate 3000 Bio compatible LC system is applied for the oxidised impurities analysis of Denosumab.The analysis was performed on a Thermo Scientific MAbPac HIC 20 column. Please Note: Blank normalized chromatogram is shown in representative profile.

Product related impurities analysis of etanercept using HIC butyl column.

Instrument Type: HPLC (Biocompatible)

Thermo Scientific ultimate 3000 BioLC system is applied for the impurities analysis of etanercept. Analysis was performed on Thermo MAbPac HIC butyl column.

Separation of related impurities of Romiplostim by Reverse Phase chromatography using wide pore phenyl column.

Instrument Type: HPLC (Biocompatible)

The Thermo Scientific Dionex UltiMate 3000 LC Bio-LC system equipped with column pre-heater is applied for the Reverse phase analysis of Romiplostim. The separation was performed on a Thermo Scientific MAbPac RP column with a UV detection at 280 nm.

AN73063: Improved Profiling of Sialylated N-Linked Glycans by Ion Chromatography-Orbitrap Mass Spectrometry

Instrument Type: ICMS

The methods proposed here use a Thermo Scientific™ Dionex™ CarboPac™ PA200 column for glycan separation by HPAE. After the separation the glycans pass through a desalting device and in to the mass spectrometer. Fragmentation of glycans in the negative mode by higher-energy collisional dissociation (HCD) provides information-rich MS2 spectra which enables glycan structure annotation. This allowed correlation of glycan structure with observed elution behavior. Here we show that in some cases, changed elution conditions can be used to resolve different glycan structures.

HPLC charged aerosol detector analysis of polysorbate 80 and insulin in a biopharmaceutical formulation

Instrument Type: HPLC-CAD

This application shows a method for HPLC-CAD analysis of polysorbate 80 and insulin in a biopharmaceutical formulation. The characterization and quantification of polysorbates is difficult because these compounds are heterogeneous mixtures with no chromophore. Because polysorbates are used in final formulations, determination of lot-to-lot variability is critical to both the pharmaceutical and biopharmaceutical industries.

CarboPac PA1-4mm Column QAR method

Instrument Type: IC

Before running any samples, Thermo Scientific recommends that you first confirm the performance of the column by reproducing the lot validation report chromatogram shipped with column. Compare your results with the one reported in the quality assurance report. At least three injections should be made. This record provides an eWorkflow for executing the QAR method on an ICS-5000+ system. An ICS-6000 can be used for this application.

AN72914: HPAE-PAD Profiling of N-linked Oligosaccharides from Glycoproteins using Dual Eluent Generation Cartridges - Gradient for Glycans from AGP

Instrument Type: IC

Glycoprotein characterization and glycosylation profiling are important tasks in the development and production of biopharmaceutical proteins. This application note demonstrates Dual EGC capability and performance for profiling N-linked oligosaccharides released from glycoproteins using high-performance anion exchange chromatography with pulsed amperometric detection (HPAE-PAD). This record shows the separation of N-linked glycans from human acid-1 glycoprotein.

AN72914: HPAE-PAD Profiling of N-linked Oligosaccharides from Glycoproteins using Dual Eluent Generation Cartridges - Gradient for Glycans from IgG

Instrument Type: IC

Glycoprotein characterization and glycosylation profiling are important tasks in the development and production of biopharmaceutical proteins. This application note demonstrates Dual EGC capability and performance for profiling N-linked oligosaccharides released from glycoproteins using high-performance anion exchange chromatography with pulsed amperometric detection (HPAE-PAD). This record shows the separation for IgG N-linked glycans.

Application of HPLC-UV-CAD in the consistency evaluation of cefepime injections

Instrument Type: HPLC-CAD

HPLC-UV-CAD method was established in this paper and the consistency of the generic drug for cefepime of injection was evaluated from the impurity profile and excipients. On the basis of UV detection and the charged aerosol detection (CAD) combined with a mixed matrix column Trinity-P1. By adjusting the organic phase and salt concentration, a method for determining the arginine of the auxiliary material was established. The determination can be made simultaneously with the main component of the drug and the salt forming ion.

Dionex CarboPac PA1 2 mm Column Performance Test Using its QAR Method

Instrument Type: IC

Before running any samples, Thermo Scientific recommends that you first confirm the performance of the column by reproducing the lot validation report chromatogram shipped with column. Compare your results with the one reported in the quality assurance report. At least three injections should be made. This record provides an eWorkflow for executing the QAR method on an ICS-5000+ system. An ICS-6000 can be used for this application.

A multi-detector platform comprising UV/Vis, charged aerosol, and single quadrupole mass spectrometric detection for comprehensive sample analysis

Instrument Type: LCMS

This application demonstrates reliable verification and quantification of the presence of extraneous compounds in a sample, such as impurities, degradation products or extractables and leachables.

AU72829: HPAE-PAD Analysis of N-linked Glycans: Improving Glycan Resolution

Instrument Type: IC

This application update demonstrates improved resolution of sialylated N-glycans on Thermo Scientific™ Dionex™ CarboPac™ PA200 columns. Starting with a recently described method, changes to elution conditions were tested to improve resolution of sialylated N-glycans enzymatically released from four different glycoproteins. Separations were first evaluated on the analytical format (3 × 250 mm column). Next, the possibility that a shorter column, such as a guard column, would allow significantly improved throughput was evaluated. Finally, a narrow bore format 1 × 250 mm column was tested.

Impurity profiling of the synthetic peptide LL-37 using high-performance liquid chromatography with combined UV and single quadrupole mass spectrometric detection

Instrument Type: LCMS

Leveraging UV detection for the determination of the impurity levels and mass detection for identification of the active pharmaceutical ingredient (API) and other product related impurities using a rapid method for synthetic peptide impurity profiling

Rapid quantitation of therapeutic antibodies in animal plasma

Instrument Type: LCMS

To demonstrate the rapid quantitation of a low-level human IgG in animal plasma by LC/MS/MS using the Thermo Scientific™ SMART Digest™ Immunoaffinity (IA) kits (including magnetic and non-magnetic versions of the Streptavidin kit, Protein A kit, and Protein G kit), which combine the IA capture and digestion process into a single well. The assay is required to be both selective and accurate.

Accurate and precise quantification of mAb-released N-glycans with an amide HILIC column

Instrument Type: UHPLC

This application note demonstrated the robustness, reproducibility, accuracy, and precision of quantification of IgG N-glycans using the Thermo Scientific™ Vanquish™ Horizon UHPLC integrated biocompatible system and Thermo Scientific™ Accucore™ 150 Amide HILIC column.

NISTmAb characterization with a high-performance RP chromatography column

Instrument Type: UHPLC

To demonstrate the robustness, reproducibility, carry-over, recovery, accuracy, and precision of quantification of the NIST monoclonal antibody IgG1K (NISTmAb) using the Thermo Scientific™ Vanquish™ Horizon UHPLC integrated biocompatible system and Thermo Scientific™ MAbPac™ RP LC columns

AB72720: A Glycopeptide Standard for Improving Glycoprotein Hydrolysis Reaction Yield for Accurate Monosaccharide Content Determination

Instrument Type: IC

This study demonstrates use of a commercially available glycopeptide standard as a control to determine glycoprotein hydrolysis reaction efficiency. Because the monosaccharide composition of glycopeptide is known, hydrolysis efficiency under a set of conditions can be easily calculated. The monosaccharide yield for the glycopeptide was compared with that of two glycoproteins under identical conditions. This comparison revealed efficient hydrolysis using the conditions tested here. Minimally the glycopeptide standard serves as a positive control for glycoprotein monosaccharide determination.

Simple charge variant profile comparison of an innovator monoclonal antibody and a biosimilar candidate

Instrument Type: UHPLC

To demonstrate the effectiveness of a simple pH gradient/ion-exchange chromatography workflow approach to the characterization of charge variant profiles of an innovator molecule (cetuximab) and a candidate biosimilar. To show the assay is simple, reproducible, easily optimized and resolves variants effectively.

Evaluation and application of salt- and pH-based ion-exchange chromatography gradients for analysis of therapeutic monoclonal antibodies

Instrument Type: UHPLC

To demonstrate the CX-1 pH buffer system is capable of charge variant determination for the majority of therapeutic monoclonal antibody species. To show the ease of optimization and improved reproducibility of this buffer system when compared to salt-based gradient systems used for charge variant analysis.

1 mm Dionex CarboPac PA200 column Performance Test Using its QAR Method with Dual Eluent Generator Cartridge Mode

Instrument Type: IC

Before running any samples, Thermo Scientific recommends that you first confirm the performance of the column by reproducing the lot validation report chromatogram shipped with column. Compare your results with the one reported in the quality assurance report. At least three injections should be made. This record provides an eWorkflow for executing the QAR method on an ICS-6000 system using Dual EGC mode.

Analysis of trace elements in Traditional Chinese Medicine (TCM)

Instrument Type: ICP-OES

This application note demonstrates the ability of the Thermo Scientific iCAP 7000 Plus Series ICP-OES to determine trace elements in materials of plant origin that are used for health benefits.

QAR Method for the CarboPac PA20-Fast-4µm, 4x100 mm Column

Instrument Type: IC

This record describes method for reproduction of column Quality Assurance Report (QAR). Analyzing a standard sample using method conditions described in QAR is recommended in order to validate column performance prior to use. This record also includes a one-click eWorkflow for executing the QAR method on a ICS 5000+ system.

AN72580: An Improved HPAE-PAD Method for Glycoprotein Monosaccharide Determination

Instrument Type: IC

This work demonstrates an improved HPAE-PAD method using a recently introduced Thermo Scientific™ Dionex™ CarboPac™ PA20 column with 4 μm particle size to simultaneously quantify six major monosaccharides present in glycoprotein acid digests. The total run time is 20 min as compared to 32 min with the method using a 6.5 μm particle size column. The smaller particle size of this column offers higher peak efficiencies, leading to high-resolution separations and allowing a shorter column format and, ultimately, significantly shorter run times. An ICS-6000 can be used for this application.

TN72478: Instrument Configuration for Native N-linked Oligosaccharide Characterization by HPAE-PAD/MS - Gradient 3

Instrument Type: ICMS

The biopharmaceutical industry requires reliable high throughput analysis of N-linked glycans. Many of the commonly used analytical approaches are based on derivatization, which is known to introduce differential loss of sialic acid and incomplete labeling. Preparation of native glycans is not subject to these concerns. This approach starts with a fast in-solution glycoprotein digestion to obtain oligosaccharides. These native glycans are then analyzed by high-performance anion-exchange chromatography with pulsed amperometric detection coupled to MS. This record is for neutral and IgG glycans.

TN72478: Instrument Configuration for Native N-linked Oligosaccharide Characterization by HPAE-PAD/MS - Gradient 2

Instrument Type: ICMS

The biopharmaceutical industry requires reliable high throughput analysis of N-linked glycans. Many of the commonly used analytical approaches are based on derivatization, which is known to introduce differential loss of sialic acid and incomplete labeling. Preparation of native glycans is not subject to these concerns. This approach starts with a fast in-solution glycoprotein digestion to obtain oligosaccharides. These native glycans are then analyzed by high-performance anion-exchange chromatography with pulsed amperometric detection coupled to MS. This record is for charged glycans.

TN72478: Instrument Configuration for Native N-linked Oligosaccharide Characterization by HPAE-PAD/MS - Gradient 1

Instrument Type: ICMS

The biopharmaceutical industry requires reliable high throughput analysis of N-linked glycans. Many of the commonly used analytical approaches are based on derivatization, which is known to introduce differential loss of sialic acid and incomplete labeling. Preparation of native glycans is not subject to these concerns. This approach starts with a fast in-solution glycoprotein digestion to obtain oligosaccharides. These native glycans are then analyzed by high-performance anion-exchange chromatography with pulsed amperometric detection coupled to MS. This record is for IgG glycans.

HILIC Separation of Glycopeptides

Instrument Type: LCMS

a UHPLC HILIC solution for the separation of glycosylated peptides following SMART digestion of a protein sample

SEC Protein aggregate analysis

Instrument Type: UHPLC

A UHPLC workflow for mAb aggregate analysis that operates continuously for weeks.

A highly sensitive high-performance liquid chromatography-charged aerosol detection method for the quantitative analysis of polysorbate 80 in protein solution

Instrument Type: HPLC-CAD

To describe the development of a highly sensitive charged aerosol detection (CAD) method for the determination of polysorbate 80, also known as Tween™ 80, in biopharmaceutical products.

High-precision, automated peptide mapping of proteins

Instrument Type: UHPLC

The biopharmaceutical industry continues to develop protein-based biotherapeutics in increasing numbers. Due to their complexity and biotechnological production, there are many attributes that need to be analyzed to guarantee their safety and efficacy. Peptide mapping is used to measure several critical quality attributes (CQA) required for the characterization of any biotherapeutic protein. The analysis is used to confirm that the correct sequence has been expressed for the protein and to check for post-translational and chemical modifications.

TN72264: HPAE-PAD N-linked Oligosaccharide Profiling of IgG

Instrument Type: IC

This technical note reports an HPAE-PAD method to profile IgG N-linked oligosaccharides and N-linked high-mannose type oligosaccharides that are atypical for human polyclonal IgG but sometimes found on mAbs. This HPAE-PAD method is orthogonal to other oligosaccharide methods applied to IgG. More information on the development and application of the separation described in the technical note can be found in Rohrer, et al. as referenced in the document.

Separation of oxidized and deamidated impurities of Infliximab by reverse phase chromatography

Instrument Type: HPLC (Biocompatible)

The Thermo Scientific Dionex UltiMate 3000 Bio-LC system is applied for the Reverse phase analysis of Infliximab. The separation was performed on a Thermo Scientific MabPac RP column with UV detection at 214 nm. All oxidized and deamidated impurities are well separated.

TN72225: Glycoprotein Monosaccharide Analysis using HPAE-PAD with Manually Prepared Eluent

Instrument Type: IC

This work describes an HPAE-PAD method for monosaccharide composition analysis using manually prepared eluent. Monosaccharide analysis using electrolytically generated eluent has been described before. Here, three commercially available proteins, IgG, fetuin, and alpha-1-acid glycoprotein, were individually subjected to two hydrolysis conditions using 1)HCl, for the amino sugars galactosamine and glucosamine, and 2)TFA, for the neutral sugars mannose, glucose, and galactose. Results for method linearity, robustness, and accuracy are presented here. An ICS-6000 can be used for this application.

High resolution charge variants analysis of recombinant human IgG1K NIST mAb

Instrument Type: HPLC-CAD

The Thermo Scientific Vanquish Flex UHPLC system with UV detection is applied for the charge variants analysis of NISTmAb Humanized IgG1K monoclonal antibody using strong cation exchange-based chromatographic separation on a MAbPac SCX-10 column. The use of Thermo Scientific CX-1 pH gradient buffers enabled high resolution separation of reference material monoclonal antibody charge variants using a 30 minute gradient.

A pre-concentration and on-line solid phase extraction setup for the LC-MS analysis of therapeutic protein mixtures

Instrument Type: LCMS

This study shows a fully automated UHPLC setup applying a simple and efficient online solid phase extraction method. Showcase assay performance by using a column cartridge for sample trapping and a Thermo Scientific™ MAbPac™ RP column for separation with water/acetonitrile-based gradients. Leverage the new technologies such as the Thermo Scientific™ Vanquish™ UHPLC platform in combination with the Thermo Scientific™ Q Exactive™ HF Hybrid Quadrupole Orbitrap mass spectrometer.

Ultrafast hydrophilic interaction UHPLC analysis of 2AA labelled N-glycans from monoclonal antibodies (mAbs)

Instrument Type: UHPLC

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.

SMART Digest compared to classic in-solution digestion of rituximab for in-depth peptide mapping characterization

Instrument Type: LCMS

This study compares the newly developed Thermo Scientific™ SMART Digest™ kit to classic in-solution protein digestion methods, focusing on protein sequence coverage and identified post-translational modifications (PTMs), including deamidation, oxidation, and glycosylation. A Thermo Scientific™ Acclaim™ VANQUISH™ C18 column with conventional water/acetonitrile-based gradients and the Thermo Scientific™ Vanquish™ Flex UHPLC system were used for separation in combination with the Thermo Scientific™ Q Exactive™ HF Hybrid Quadrupole-Orbitrap™ mass spectrometer.

Tandem UHPLC operation for high-throughput LC-MS peptide mapping analyses

Instrument Type: LCMS

To demonstrate the use of a multi-pump UHPLC system and enable tandem analysis with two columns in parallel, addressing productivity and throughput improvement of existing LC-MS methods.