A5043224453- Protein purification and stability
- Monoclonal and Polyclonal Antibodies Research
- Protein Structure and Dynamics
- Viral Infectious Diseases and Gene Expression in Insects
- Mass Spectrometry Techniques and Applications
- Transgenic Plants and Applications
- Analytical Chemistry and Chromatography
- Microfluidic and Capillary Electrophoresis Applications
- Respiratory viral infections research
- Bacteriophages and microbial interactions
- Protein Interaction Studies and Fluorescence Analysis
- Enzyme Structure and Function
- Viral gastroenteritis research and epidemiology
- Escherichia coli research studies
- Drug Solubulity and Delivery Systems
- Intramuscular injections and effects
- Neuropeptides and Animal Physiology
- Botulinum Toxin and Related Neurological Disorders
- Pneumonia and Respiratory Infections
- Signaling Pathways in Disease
- Mesenchymal stem cell research
- Biosimilars and Bioanalytical Methods
- RNA Research and Splicing
- Oral health in cancer treatment
- 3D Printing in Biomedical Research
AstraZeneca (United States)
2019-2024
Baqiyatallah University of Medical Sciences
2023
University of Kansas
2008-2022
Pharmaceutical Formulations (United States)
2018
Altimmune (United States)
2017
Concentration-dependent reversible self-association (RSA) of monoclonal antibodies (mAbs) poses a challenge to their pharmaceutical development as viable candidates for subcutaneous delivery. While the role antigen-binding fragment (Fab) in initiating RSA is well-established, little evidence supports involvement crystallizable (Fc). In this report, variety biophysical tools, including hydrogen exchange mass spectrometry, are used elucidate protein interface such non-covalent protein-protein...
There is a need for new analytical approaches to better characterize the nature of concentration-dependent, reversible self-association (RSA) monoclonal antibodies (mAbs) directly, and with high resolution, when these proteins are formulated as highly concentrated solutions. In work reported here, hydrogen exchange mass spectrometry (HX-MS) was used define concentration-dependent RSA interface, effects association on backbone dynamics an IgG1 mAb (mAb-C). Dynamic light scattering, chemical...
This study compares the local conformational dynamics and physical stability of an IgG1 mAb (mAb-A) with its corresponding YTE (M255Y/S257T/T259E) mutant (mAb-E), which was engineered for extended half-life in vivo. Structural measured using hydrogen/deuterium (H/D) exchange mass spectrometry while protein differential scanning calorimetry (DSC) size exclusion chromatography (SEC). The mutation induced differences H/D kinetics at both pH 6.0 7.4. Segments covering sites FcRn binding epitopes...
Monoclonal antibodies (mAbs) are complex molecular structures. They often prone to development challenges particularly at high concentrations due undesired solution properties such as reversible self-association, viscosity, and liquid–liquid phase separation. In addition formulation optimization, applying protein engineering can provide an alternative mitigation strategy. Protein during the discovery great benefits optimize properties, resulting in improved developability profiles. Here, we...
Undesired solution behaviors such as reversible self-association (RSA), high viscosity, and liquid-liquid phase separation can introduce substantial challenges during development of monoclonal antibody formulations. Although a global mechanistic understanding RSA (i.e., native protein-protein interactions) is sufficient to develop robust formulation controls, its mitigation via protein engineering requires knowledge the sites interactions. In study reported here, we coupled our previous...
Electrostatically driven attractions between proteins can result in issues for therapeutic protein formulations such as solubility limits, aggregation, and high solution viscosity. Previous work showed that a model monoclonal antibody displayed large potentially problematic electrostatically at typical pH (5-8) ionic strength conditions (∼10-100 mM). Molecular simulations of hybrid coarse-grained (1bC/D, one bead per charged site domain) were used to predict potential point mutations...
Proteins display a broad peak in 250-300 nm region of their UV spectrum containing multiple overlapping bands arising from the aromatic rings phenylalanine, tyrosine, and tryptophan residues. Employing high resolution 2(nd) derivative absorbance spectroscopy, these absorption can be highly resolved therefore provide very sensitive measure changes local microenvironment side chains. This has traditionally been used to detect both subtle dramatic (i.e., unfolding) conformational alterations...
The influence of PEGylation on the thermal stability small therapeutic proteins was evaluated using two model proteins. Changes in midpoint unfolding and ability to properly refold after denaturation were monitored by differential scanning calorimetry (DSC) as a function pH. results showed that increased both well their denaturation. DSC compared traditional accelerated data collected size exclusion high performance liquid chromatography (SE-HPLC). agreed reasonably with those from SE-HPLC...
Increased protein solubility is known to correlate with an increase in the proportion of lysine over arginine residues. Previous work has shown that aggregation propensity a single-chain variable fragment (scFv) does not its conformational stability or native-state protein–protein interactions. Here, we test hypothesis driven by colloidal partially unfolded states, studying behavior scFv mutants harboring single multiple site-specific mutations denaturing buffers. In 6 M guanidine...
Monoclonal antibodies are a class of biotherapeutics used for an increasing variety disorders, including cancer, autoimmune, neurodegenerative, and viral diseases. Besides their antigen specificity, therapeutic use also mandates control solution interactions colloidal properties in order to achieve stable, efficacious, non-immunogenic, low viscosity antibody at concentrations the range 50–150 mg/mL. This requires characterization reversible self-association, aggregation, weak attractive...
A barrier to the use of hydrogen exchange-mass spectrometry (HX-MS) in many contexts, especially analytical characterization various protein therapeutic candidates, is that differences temperature, pH, ionic strength, buffering agent, or other additives can alter chemical exchange rates, making HX data gathered under differing solution conditions difficult compare. Here, we present demonstrating rates be substantially altered not only by well-established variables temperature and pH but also...
Light exposure of a monoclonal antibody formulation containing polysorbate 80 (PS80) leads to cis/trans isomerization monounsaturated and polyunsaturated fatty acids. This was monitored by positive electrospray ionization mass spectrometry intact PS80 components as well negative ion analysis free acids generated via esterase-catalyzed hydrolysis. The light-induced unsaturated in required the presence antibody, or, at minimum (for mechanistic studies), combination N-acetyltryptophan amide...