A5089993749- Protein purification and stability
- Monoclonal and Polyclonal Antibodies Research
- HER2/EGFR in Cancer Research
- Mass Spectrometry Techniques and Applications
- Advanced Proteomics Techniques and Applications
- Maternal and Perinatal Health Interventions
- Bacterial Identification and Susceptibility Testing
- Biosimilars and Bioanalytical Methods
- Advanced biosensing and bioanalysis techniques
- Biosensors and Analytical Detection
- Palliative Care and End-of-Life Issues
- Maternal Mental Health During Pregnancy and Postpartum
- Metabolomics and Mass Spectrometry Studies
- Glycosylation and Glycoproteins Research
- Advanced Biosensing Techniques and Applications
- Maternal and fetal healthcare
- Family and Patient Care in Intensive Care Units
University of Groningen
2019-2023
Manchester Royal Infirmary
2021
Obafemi Awolowo University Teaching Hospitals Complex
2008
Cation-exchange chromatography is a widely used approach to study charge heterogeneity of monoclonal antibodies. Heterogeneity may arise both in vitro and vivo because the susceptibility antibodies undergo chemical modifications. Modifications adversely affect potency drug, induce immunogenicity or pharmacokinetics. In this study, we evaluated application optimized pH gradient systems for separation variants trastuzumab after forced degradation study. gradient-based elution resulted...
Pertuzumab is a monoclonal antibody used for the treatment of HER2-positive breast cancer in combination with trastuzumab. Charge variants trastuzumab have been extensively described literature; however, little known about charge heterogeneity pertuzumab. Here, changes ion-exchange profile pertuzumab were evaluated by pH gradient cation-exchange chromatography after stressing it up to 3 weeks at physiological and elevated 37 °C. Isolated arising under stress conditions characterized peptide...
Hydrogen-deuterium exchange mass spectrometry (HDX-MS) is a method to probe the solvent accessibility and conformational dynamics of protein or protein-ligand complex with respect exchangeable amide hydrogens. Here, we present application HDX-MS determine binding sites Affimer reagents monoclonal antibodies trastuzumab pertuzumab, respectively. Intact subunit level analysis antibody-affimer complexes showed significant protection from HDX in antibody Fab region upon affimer binding....
Asparagine deamidation and aspartic acid isomerization in the complementarity determining regions (CDRs) of monoclonal antibodies may alter their affinity to target antigen. Trastuzumab has two hot spots for one position CDRs. Little is known how complex formation with its antigen HER2 affects these modifications. Modifications CDRs trastuzumab were thus compared between free antibody trastuzumab-HER2 when stressed under physiological conditions at 37°C. Complex stability upon stressing...
Trastuzumab and pertuzumab are monoclonal antibodies used in the treatment of human epidermal growth factor receptor-2 (HER2)-positive breast cancer. Therapeutic proteins may undergo chemical modifications that affect results bioanalytical assays, as well their therapeutic efficacy. Modifications arise during production storage, after administration to patients. Studying vivo biotransformation monoclonal, requires enrichment from plasma discriminate them endogenous antibodies, other...
Trastuzumab is known to be heterogeneous in terms of charge. Stressing trastuzumab under physiological conditions (pH 7.4 and 37 °C) increases charge heterogeneity further. Separation variants stressed at the intact protein level challenging due increasing complexity making it difficult obtain pure for further characterization. Here we report an approach revealing subunit by pH gradient cation-exchange chromatography. subunits were generated after limited proteolytic cleavage with papain,...
Therapeutic proteins (TPs) are known to be heterogeneous due modifications that occur during the production process and storage. Modifications may also in TPs after their administration patients vivo biotransformation. Ligand binding assays, which widely used bioanalysis of body fluids, typically unable distinguish such modifications. Liquid chromatography coupled mass spectrometry is being increasingly study TPs, but its use biotransformation has been limited until now. We present a novel...
Intact Transition Epitope Mapping—One-step Non-covalent force Exploitation (ITEM-ONE) analysis reveals an assembled epitope on the surface of Pertuzumab, which is recognized by anti-Pertuzumab affimer 00557_709097. It encompasses amino acid residues NSGGSIYNQRFKGR, are part CDR2, as well FTLSVDR, located variable region Pertuzumab’s heavy chain and together form a area 1381.46 Å2. Despite not being partial sequence FTLSVDR marks unique proteotypic Pertuzumab peptide. Binding between intact...
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Abstract Aim The unique situation created by the global pandemic has led us to re-evaluate how we disseminate information about service provision patients and their relatives. aim of this project is create a patient centred video, explaining Major Trauma (MT) service. Methodology Participants who accessed MT from May 2020- July 2020 were approached evaluate thoughts on through open questions. raw data was analysed using thematic analysis. themes generated incentivizing force behind creation...