Ionic liquid-based strategy for predicting protein aggregation propensity and thermodynamic stability
<p>Novel drug candidates are continuously being developed to combat the most life-threatening diseases; however, many promising protein therapeutics are dropped from the pipeline. During biological and industrial processes, protein therapeutics are exposed to various stresses such as fluctuati...
| Main Authors: | , , , |
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| Format: | Journal article |
| Language: | English |
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American Chemical Society
2022
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| _version_ | 1826310131252461568 |
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| author | Shmool, TA Martin, LK Matthews, RP Hallett, JP |
| author_facet | Shmool, TA Martin, LK Matthews, RP Hallett, JP |
| author_sort | Shmool, TA |
| collection | OXFORD |
| description | <p>Novel drug candidates are continuously being developed to combat the most life-threatening diseases; however, many promising protein therapeutics are dropped from the pipeline. During biological and industrial processes, protein therapeutics are exposed to various stresses such as fluctuations in temperature, solvent pH, and ionic strength. These can lead to enhanced protein aggregation propensity, one of the greatest challenges in drug development. Recently, ionic liquids (ILs), in particular, biocompatible choline chloride ([Cho]Cl)-based ILs, have been used to hinder stress-induced protein conformational changes. Herein, we develop an IL-based strategy to predict protein aggregation propensity and thermodynamic stability. We examine three key variables influencing protein misfolding: pH, ionic strength, and temperature. Using dynamic light scattering, zeta potential, and variable temperature circular dichroism measurements, we systematically evaluate the structural, thermal, and thermodynamic stability of fresh immunoglobin G4 (IgG4) antibody in water and 10, 30, and 50 wt % [Cho]Cl. Additionally, we conduct molecular dynamics simulations to examine IgG4 aggregation propensity in each system and the relative favorability of different [Cho]Cl-IgG4 packing interactions. We re-evaluate each system following 365 days of storage at 4 °C and demonstrate how to predict the thermodynamic properties and protein aggregation propensity over extended storage, even under stress conditions. We find that increasing [Cho]Cl concentration reduced IgG4 aggregation propensity both fresh and following 365 days of storage and demonstrate the potential of using our predictive IL-based strategy and formulations to radically increase protein stability and storage.</p> |
| first_indexed | 2024-03-07T07:47:36Z |
| format | Journal article |
| id | oxford-uuid:d9a5646e-9fad-434c-a05f-63a948d4d125 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T07:47:36Z |
| publishDate | 2022 |
| publisher | American Chemical Society |
| record_format | dspace |
| spelling | oxford-uuid:d9a5646e-9fad-434c-a05f-63a948d4d1252023-06-15T12:47:37ZIonic liquid-based strategy for predicting protein aggregation propensity and thermodynamic stabilityJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:d9a5646e-9fad-434c-a05f-63a948d4d125EnglishSymplectic ElementsAmerican Chemical Society 2022Shmool, TAMartin, LKMatthews, RPHallett, JP<p>Novel drug candidates are continuously being developed to combat the most life-threatening diseases; however, many promising protein therapeutics are dropped from the pipeline. During biological and industrial processes, protein therapeutics are exposed to various stresses such as fluctuations in temperature, solvent pH, and ionic strength. These can lead to enhanced protein aggregation propensity, one of the greatest challenges in drug development. Recently, ionic liquids (ILs), in particular, biocompatible choline chloride ([Cho]Cl)-based ILs, have been used to hinder stress-induced protein conformational changes. Herein, we develop an IL-based strategy to predict protein aggregation propensity and thermodynamic stability. We examine three key variables influencing protein misfolding: pH, ionic strength, and temperature. Using dynamic light scattering, zeta potential, and variable temperature circular dichroism measurements, we systematically evaluate the structural, thermal, and thermodynamic stability of fresh immunoglobin G4 (IgG4) antibody in water and 10, 30, and 50 wt % [Cho]Cl. Additionally, we conduct molecular dynamics simulations to examine IgG4 aggregation propensity in each system and the relative favorability of different [Cho]Cl-IgG4 packing interactions. We re-evaluate each system following 365 days of storage at 4 °C and demonstrate how to predict the thermodynamic properties and protein aggregation propensity over extended storage, even under stress conditions. We find that increasing [Cho]Cl concentration reduced IgG4 aggregation propensity both fresh and following 365 days of storage and demonstrate the potential of using our predictive IL-based strategy and formulations to radically increase protein stability and storage.</p> |
| spellingShingle | Shmool, TA Martin, LK Matthews, RP Hallett, JP Ionic liquid-based strategy for predicting protein aggregation propensity and thermodynamic stability |
| title | Ionic liquid-based strategy for predicting protein aggregation propensity and thermodynamic stability |
| title_full | Ionic liquid-based strategy for predicting protein aggregation propensity and thermodynamic stability |
| title_fullStr | Ionic liquid-based strategy for predicting protein aggregation propensity and thermodynamic stability |
| title_full_unstemmed | Ionic liquid-based strategy for predicting protein aggregation propensity and thermodynamic stability |
| title_short | Ionic liquid-based strategy for predicting protein aggregation propensity and thermodynamic stability |
| title_sort | ionic liquid based strategy for predicting protein aggregation propensity and thermodynamic stability |
| work_keys_str_mv | AT shmoolta ionicliquidbasedstrategyforpredictingproteinaggregationpropensityandthermodynamicstability AT martinlk ionicliquidbasedstrategyforpredictingproteinaggregationpropensityandthermodynamicstability AT matthewsrp ionicliquidbasedstrategyforpredictingproteinaggregationpropensityandthermodynamicstability AT hallettjp ionicliquidbasedstrategyforpredictingproteinaggregationpropensityandthermodynamicstability |