Article
Medicine, Research & Experimental
Victoria E. Wood, Kate Groves, Lok Man Wong, Luyan Kong, Christopher Bird, Meenu Wadhwa, Milena Quaglia, Paul Matejtschuk, Paul A. Dalby
Summary: Protein engineering and formulation for therapeutic proteins present a major challenge in the biopharmaceutical industry due to the need for prolonged shelf-life. Understanding the impact of mutations and formulations on protein structure and dynamics can lead to more predictive approaches for improvement. Studies using intrinsic fluorescence analysis and hydrogen deuterium exchange mass spectrometry have revealed insights into the reorganization and stabilization of specific protein loops, providing valuable information for enhancing protein stability and reducing aggregation rates.
MOLECULAR PHARMACEUTICS
(2022)
Article
Biochemistry & Molecular Biology
Joelle A. J. Housmans, Guiqin Wu, Joost Schymkowitz, Frederic Rousseau
Summary: Disruptions or instabilities in protein folding can lead to the accumulation of misfolded proteins and the formation of protein aggregates, which are associated with human diseases and pose challenges in biopharmaceutical manufacturing. However, protein aggregates can also be functional and have applications in biomaterials, therapeutics, and food improvement. Understanding the steps involved in protein aggregation is crucial for developing diagnostic methods, treatments, and applications in various fields. The study of protein aggregation is challenging due to its complex and dynamic nature.
Article
Biochemistry & Molecular Biology
Cheng Zhang, Nuria Codina, Jiazhi Tang, Haoran Yu, Nesrine Chakroun, Frank Kozielski, Paul A. Dalby
Summary: The research findings highlighted the different destabilizing pathways of protein structure under low pH and high temperature stress conditions, revealing the potential for stabilizing mutations in the CL domain and C-L-C(H)1 interface. The analysis also identified key salt bridge losses as a cause of conformational changes under different stresses, with implications for future engineering efforts to enhance stability.
COMPUTATIONAL AND STRUCTURAL BIOTECHNOLOGY JOURNAL
(2021)
Article
Biochemistry & Molecular Biology
Jingbo Liu, Hongyu Jiang, Min Zhang, Ping Gong, Meng Yang, Ting Zhang, Xuanting Liu
Summary: This study evaluated the structure of ions-regulated gelation of egg white protein through aggregation kinetics model, showing that the addition of Na2SO4 significantly influenced the gel structure, leading to improved gel network strength and porosity.
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
(2022)
Article
Biochemistry & Molecular Biology
Javed Masood Khan, Ajamaluddin Malik, Prerna Sharma, Sadaf Fatima
Summary: In this study, the impact of the negatively charged detergent sodium dodecyl sulfate (SDS) on insulin amyloid fibrillation was investigated. It was found that SDS can accelerate insulin aggregation at pH 2.0, but not at pH 7.4.
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
(2023)
Review
Pharmacology & Pharmacy
Harini Narayanan, Fabian Dingfelder, Alessandro Butte, Nikolai Lorenzen, Michael Sokolov, Paolo Arosio
Summary: Successful biologics must meet requirements of both activity and physicochemical features, which can be optimized with the help of artificial intelligence and machine learning to reduce development time and costs. Machine learning is currently applied in biologics discovery and development, particularly in areas such as protein engineering, early biophysical screening, and formulation.
TRENDS IN PHARMACOLOGICAL SCIENCES
(2021)
Article
Biochemistry & Molecular Biology
Qinqin Cheng, Xiao-Nan Zhang, Lei Zhang, Jingwen Chen, Yiling Wang, Yong Zhang
Summary: This study generated PAR polymer-GCSF conjugates through the combination of PAR polymers and human granulocyte colony-stimulating factor (GCSF) protein, which demonstrated excellent in vivo efficacy and have the potential to be used as carriers for therapeutic proteins or peptides with improved pharmacological activities.
Article
Multidisciplinary Sciences
Emanuele Scalone, Luca Broggini, Cristina Visentin, Davide Erba, Fran Bacic Toplek, Kaliroi Peqini, Sara Pellegrino, Stefano Ricagno, Cristina Paissoni, Carlo Camilloni
Summary: Understanding the mechanisms of protein aggregation is crucial for treating over 50 incurable diseases. A new hybrid structure model presented in this study effectively captures the essential structural and kinetic aspects of protein aggregation through molecular dynamics simulations. This model can help observe the formation of primary nuclei and the growth of fibrils, providing a structural basis for designing drugs targeting amyloidogenic diseases.
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
(2022)
Article
Chemistry, Physical
Jia-Liang Shen, Min-Yeh Tsai, Nicholas P. Schafer, Peter G. Wolynes
Summary: Protein aggregation plays a crucial role in determining the structure of membraneless organelles and the pathogenesis of diseases, but the diverse types of aggregates and inherent stochastic nature present challenges to theoretical and computational modeling. Introducing a new approach to model stochastic aggregation kinetics, which incorporates noise into averaged equations, helps to explore aggregation mechanisms and statistics more efficiently. The second stochasticization scheme reveals a scaling law that correlates fluctuations in aggregate size with the total number of monomers, demonstrating the value of bridging the gap between in vivo cell biology and detailed modeling.
JOURNAL OF PHYSICAL CHEMISTRY B
(2021)
Review
Chemistry, Medicinal
Samadhi Vitharana, Joshua M. Stillahn, Derrick S. Katayama, Charles S. Henry, Mark Cornell Manning
Summary: The field of formulation and stabilization of protein therapeutics is extensive. This review discusses the application of formulation principles to stabilize biopharmaceutical proteins during bioprocessing and manufacturing, considering each unit operation involved in drug substance preparation. The impact of the container on stability is also addressed.
JOURNAL OF PHARMACEUTICAL SCIENCES
(2023)
Article
Chemistry, Medicinal
Christina Besada, Abrar Hakami, Gayatri Pillai, Kyle Yetsko, Nhu Truong, Timothy Little, Sharon Pantano, Anil Dmello
Summary: The study aims to develop a PAL microcapsule formulation to deplete phenylalanine in the gastrointestinal tract. The results show that PAL activity decreases over time, but product inhibition can be relieved by adding bovine serum albumin. Additionally, PAL exhibits maximum activity at pH 8.5 and requires buffering to maintain activity.
JOURNAL OF PHARMACEUTICAL SCIENCES
(2022)
Article
Chemistry, Medicinal
Helen Haixia Wu, Maureen Crames, Yangjie Wei, Dongmei Liu, Kristina Gueneva-Boucheva, Ikbae Son, Lee Frego, Fei Han, Rachel Kroe-Barrett, Andrew Nixon, Michael Marlow
Summary: Monoclonal antibodies, especially IgGs and Ig-based molecules, are widely used biotherapeutic drugs. The engineering of the Fc domain of these drugs to optimize their interactions with Fc gamma receptors (FcgRs) has been extensively studied in recent years. However, limited information is available regarding the effects of mutations and isotype differences on the physicochemical properties and manufacturability of these drugs. This study systematically characterizes the effects of mutations and IgG4 isotype on the stability and manufacturability of six Fc variants, providing a basis for selecting appropriate Fc regions and predicting the impact of mutations on the drug development process.
JOURNAL OF PHARMACEUTICAL SCIENCES
(2022)
Article
Chemistry, Medicinal
Kelvin B. Rembert, Jifeng Zhang, Young Jong Lee
Summary: The impact of five representative Hofmeister salts on the thermal stability and aggregation kinetics of a slightly acidic monoclonal antibody was investigated. The results showed that MgCl2 and NaSCN significantly decreased the unfolding transition temperature of the antibody, while the nucleation rate was influenced by all salts tested, with different trends depending on the surface charge of the antibody. The observed effects can be attributed to ion-specific mechanisms dominated by short-range amide backbone binding and long-range electrostatic binding of ions to charged amino acid side chains.
JOURNAL OF PHARMACEUTICAL SCIENCES
(2023)
Article
Chemistry, Applied
Loes J. G. Hoppenreijs, Sarah E. Brune, Rebekka Biedendieck, Rainer Krull, Remko M. Boom, Julia K. Keppler
Summary: The role of intramolecular disulfide bonds in the fibrillation of beta-lactoglobulin (BLG) at 80°C and pH 2 was investigated. Removal of disulfide bonds affected the hydrolysis rate and aggregation kinetics, with complete removal leading to immediate aggregation. These findings have implications for the production of functional protein aggregates and the prevention of pathogenic fibrillation.
FOOD HYDROCOLLOIDS
(2023)
Article
Environmental Sciences
Biao Jing, Wang Wan, Bo Hu, Wenhan Jin, Zhenduo Zhang, Congcong Peng, Mengdie Wang, Jintai Deng, Xuepeng Dong, Yu Liu, Zhenming Gao
Summary: Plastic nanoparticles can deteriorate proteome stability, compromise cellular protein homeostasis, and cause global proteome misfolding and aggregation, leading to negative impacts on cellular fitness.
ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY
(2023)
Article
Pharmacology & Pharmacy
Andrea Arsiccio, Paul Matejtschuk, Ernest Ezeajughi, Andrew Riches-Duit, Anwen Bullen, Kiran Malik, Sanj Raut, Roberto Pisano
INTERNATIONAL JOURNAL OF PHARMACEUTICS
(2020)
Article
Pharmacology & Pharmacy
Arnold Duralliu, Paul Matejtschuk, Paul Stickings, Laura Hassall, Robert Tierney, Daryl R. Williams
Article
Biochemistry & Molecular Biology
Henry C. Wilkinson, Paul A. Dalby
Summary: This study explores the effects of site-specific saturation mutagenesis within enzyme active sites and the potential benefits of incorporating noncanonical amino acids (ncAA) on improving enzyme activity and stability simultaneously. By fine-tuning residue 385 with different aromatic ring substituents, the researchers were able to significantly enhance specific activity, catalytic performance, and stability of the enzyme, showcasing the advantages of expanding designer libraries to include ncAAs.
Article
Biophysics
Valentina A. Spiteri, James Doutch, Robert P. Rambo, Jayesh Gor, Paul A. Dalby, Stephen J. Perkins
Summary: A comparative structural study of glycosylated and deglycosylated monoclonal human IgG1 revealed perturbations in the Fab-Fc separation after deglycosylation, with a broader conformational spectrum observed in the Fc region of deglycosylated IgG1 compared to glycosylated IgG1. This variability in Fc conformations was found to account for the loss of binding to the Fc gamma receptor in deglycosylated IgG1.
BIOPHYSICAL JOURNAL
(2021)
Article
Biochemical Research Methods
John E. Hales, Samir Aoudjane, Gabriel Aeppli, Paul A. Dalby
Summary: The adoption of process analytical technologies in the biopharmaceutical industry can reduce the cost of therapeutic drugs and facilitate investigation of new bioprocesses. Control of critical process parameters is crucial for ensuring product quality, but developing the necessary sophisticated analytical technologies has proven to be a challenge.
JOURNAL OF CHROMATOGRAPHY A
(2021)
Article
Medicine, Research & Experimental
Victoria E. Wood, Kate Groves, Lok Man Wong, Luyan Kong, Christopher Bird, Meenu Wadhwa, Milena Quaglia, Paul Matejtschuk, Paul A. Dalby
Summary: Protein engineering and formulation for therapeutic proteins present a major challenge in the biopharmaceutical industry due to the need for prolonged shelf-life. Understanding the impact of mutations and formulations on protein structure and dynamics can lead to more predictive approaches for improvement. Studies using intrinsic fluorescence analysis and hydrogen deuterium exchange mass spectrometry have revealed insights into the reorganization and stabilization of specific protein loops, providing valuable information for enhancing protein stability and reducing aggregation rates.
MOLECULAR PHARMACEUTICS
(2022)
Article
Immunology
Kai-Thomas Schneider, Toni Kirmann, Esther Veronika Wenzel, Jan-Hendrik Grosch, Saskia Polten, Doris Meier, Marlies Becker, Paul Matejtschuk, Michael Hust, Giulio Russo, Stefan Duebel
Summary: Generation of sequence defined antibodies from universal libraries by phage display has been established as a robust method. ScFv-Fcs can be effective as neutralizing antibodies against a panel of pathogens and toxins, but may have variable stability. A generic lyophilization protocol has been developed to improve their shelf life, showing preservation of initial properties without compromising binding activity after storage.
FRONTIERS IN CELLULAR AND INFECTION MICROBIOLOGY
(2021)
Article
Pharmacology & Pharmacy
Kiran P. Malik, Chinwe Duru, Paul Stickings, Esther Veronika Wenzel, Michael Hust, Paul Matejtschuk
Summary: Plastic microtubes are investigated as an alternative container for small volume fills, but issues with stability may arise when freeze-drying more labile materials. In this study, it is demonstrated that the microtube format is unsuitable for ensuring the stability of the monoclonal antibody tested.
Article
Medicine, Research & Experimental
Mark-Adam W. Kellerman, Teresa Almeida, Timothy R. Rudd, Paul Matejtschuk, Paul A. Dalby
Summary: Structure-function relationships in proteins are influenced by the trade-off between stability and bioactivity. This study uses NMR spectroscopy to identify key amino acid residues in granulocyte colony-stimulating factor (G-CSF) that affect global stability and undergo conformational changes. It is found that certain residues are important for both stability and function, suggesting a possible link between the two. These findings provide insights into the crucial role of specific residues in the structure-function trade-off of proteins.
MOLECULAR PHARMACEUTICS
(2022)
Article
Biochemistry & Molecular Biology
Paul Matejtschuk, Christopher Bird, Ernest Ezeajughi, Kirsty MacLellan-Gibson, Meenu Wadhwa
Summary: This study compares the impact of buffer choice and sodium chloride addition on the outcome of freeze-drying biologics, and finds that the addition of sodium chloride can significantly reduce residual moisture content.
FRONTIERS IN MOLECULAR BIOSCIENCES
(2022)
Article
Medicine, Research & Experimental
Hongyu Zhang, Paul A. Dalby
Summary: Combined administration of antibody therapeutics is beneficial, but the stability of the antibodies may be affected when combined into a single product.
MOLECULAR PHARMACEUTICS
(2022)
Article
Biochemistry & Molecular Biology
Cheng Zhang, Nuria Codina, Jiazhi Tang, Haoran Yu, Nesrine Chakroun, Frank Kozielski, Paul A. Dalby
Summary: The research findings highlighted the different destabilizing pathways of protein structure under low pH and high temperature stress conditions, revealing the potential for stabilizing mutations in the CL domain and C-L-C(H)1 interface. The analysis also identified key salt bridge losses as a cause of conformational changes under different stresses, with implications for future engineering efforts to enhance stability.
COMPUTATIONAL AND STRUCTURAL BIOTECHNOLOGY JOURNAL
(2021)
Article
Biochemistry & Molecular Biology
Hongyu Zhang, Yang Yang, Cheng Zhang, Suzanne S. Farid, Paul A. Dalby
Summary: By training a neural network model to predict spectra based on the temperature-dependence of intrinsic fluorescence emission under native conditions, hidden information related to stability was discovered in native fluorescence spectra. This method could potentially enable rapid screening of therapeutic protein variants and formulations based on spectroscopic measurements at non-denaturing temperatures.
COMPUTATIONAL AND STRUCTURAL BIOTECHNOLOGY JOURNAL
(2021)