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Characterization of the N-glycosylation Pattern of Antibodies
Arndt Asperger, Dirk Wunderlich, Marcus Macht, and Pierre-Olivier Schmit
This technical note describes the analysis of the N-Glycosylation pattern on antibodies using complementary mass spectrometric techniques. LC-ESI-UHR-TOF analysis provides high-resolution data for intact antibodies and released antibody chains allowing rapid assessment of major N-glycosylation isoforms. Complementary LC-MALDI-TOF/TOF then provides in-depth characterization of these N-glycopeptide patterns, generated from the digested antibodies, generating a an extremely detailed picture of the highly complex patterns of N-glycosylation. Read More |
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Edmass Verficiation of Terminal Sequences in Proteins
Anja Resemann and Detlev Suckau
This application note describes a method using the Edmass (Edman-style ladder sequencing approach) capabilities of a MALDI-TOF mass spectrometer for semi-automatic protein sequencing and automatic sequence validation work. This method can be easily applied to QC applications (such as top-down sequencing) in biotherapeutics, enabling quick and easily interpretable answers. Read More |
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N–and C–Terminal Sequence Determination of Intact Proteins
Anja Resemann and Detlev Suckau
This application note describes a procedure using the recently introduced 1,5-diaminonaphthalene (DAN) matrix and the latest MALDI-TOF/TOF instrumentation to automate the entire process of sample preparation and data acquisition to N- and C-terminal sequence assignments in intact proteins. Additional developments in dedicated top-down analysis software are also described. Read More |
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Automated Antibody Verification for Lead Discovery
Anja Resemann and Detlev Suckau
Monoclonal antibodies play an increasing role as therapeutic agents in the pharmaceutical industry, however antibody lead discovery and optimization processes require several steps. In this application note, we describe methods to automate characterization at each stage of the therapeutic pipeline. Read More |
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An Automated Solution to Characterize Intact and Digested Proteins
Christian Albers, Laura Main, Carsten Baessmann, and Wolfgang Jabs
In this application note we will apply a fully automated workflow to the QC characterization of three proteins, including intact IgG 1, digested transferrin, and digested bovine serum albumin. This workflow is a push-button solution for non-specialist operators to generate high-quality, accurate data for automatic comparison with laboratory reference standards. Read More |
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Automated UPLC–MS Analysis of Protein Sequence Variants
Ryan Hylands, Malcolm J. Saxton, Joanne Waters, Wolfgang Jabs, and Stuart Pengelley
Establishing methods for routine analysis results in a much higher throughput workflow, together with reduced user intervention and generation of consistent and reliable data processing. Here, we describe an automated UPLC–MS based method for the analysis and quality confirmation of an intact protein for protein sequence variants. The workflow implemented allows for data acquisition, processing, and analysis to occur with minimal user intervention. Read More |
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Precise Molecular Weight Determination of Intact Proteins
Anja Resemann, Rainer Paape, and Detlev Suckau
This application note describes recent advances in MALDI-TOF/TOF instrumentation to allow for higher digitization rates yielding higher quality data then was previously possible. This, in combination with a new MALDI matrix (DHAP) that enhances the formation of multiply charged ions, allows for more accurate molecule weight determination of intact proteins. The accurate intact molecular weight analysis adds an additional characterization tool to the already considerable range of biopharmaceutical characterization workflows which are possible with MALDI-TOF/TOF. Read More |
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Rapid Quality Control of Biopharmaceutical Products
Laura Main, Wolfgang Jabs, Arndt Asperger, Dirk Wunderlich, Marcus Macht, and Pierre-Olivier Schmit
Due to the fact that biological therapeutics, such as monoclonal antibodies, are usually manufactured in a living system by recombinant technology, these systems can be sensitive to minor changes in the manufacturing process. Slight modifications may adversely affect the safety and efficacy of the drug. In this application note, we describe an open access method for the rapid quality control characterization of biotherapeutic drugs that can be utilized by non-technical experts. Read More |
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Drug Metabolite Detection and Identification
George McLeod, Siân Ives, and Thomas Amos Jacobsen
Characterization of the metabolites of drug candidates is a critical step in drug development. The observed activity and toxicity of a drug molecule may be severely impacted by metabolic modification. In this application brief, we demonstrate the use of software-based metabolite prediction coupled with high-performance Qq-TOF LC–MS and component-finding algorithms for the discovery and identification of metabolites of the well-known drug Verapamil. Read More |
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Simultaneous Quantitative and Qualitative Drug Metabolism Investigations
Don Richards
The ability to simultaneously collect quantitative and qualitative information from a DMPK- analysis has the potential to significantly increase productivity in pharmaceutical drug discovery and development. This work describes an investigation using the latest quadrupole-time-of-flight mass spectrometer to obtain clearance data, metabolite identification, structure elucidation, and metabolite profiles from Cytochrome P450 microsomal incubations in a single sample set. Read More |
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