Conformational stability and self-association equilibrium in biologics

Clarkson, Benjamin R.; Schön, Arne; Freire, Ernesto

doi:10.1016/j.drudis.2015.11.007

Cited by 25 publications

(18 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The peak at 230 nm is suggestive of aggregation or disordered proteins. This disordered conformation is unstructured to partially structured, dominated by random coils and pre-molten globules, 59 and similar to intrinsically disordered proteins (IDPs) which are without a regular secondary structure or have an absence of a three-dimensional structure and is detected by CD as “random coil”, “unordered”, or “disordered”. This study may be extrapolated to some IDPs or provide some clues to diseases caused by protein misfolding.…”

Section: Discussionmentioning

confidence: 99%

Heparin Accelerates the Protein Aggregation via the Downhill Polymerization Mechanism: Multi-Spectroscopic Studies to Delineate the Implications on Proteinopathies

et al. 2021

View full text Add to dashboard Cite

Heparin is one of the members of the glycosaminoglycan (GAG) family, which has been associated with protein aggregation diseases including Alzheimer’s disease, Parkinson’s disease, and prion diseases. Here, we investigate heparin-induced aggregation of bovine serum albumin (BSA) using different spectroscopic techniques [absorption, 8-anilino-1-naphthalene sulfonic acid (ANS) and thioflavin T (ThT) fluorescence binding, and far- and near-UV circular dichroism]. Kinetic measurements revealed that heparin is involved in the significant enhancement of aggregation of BSA. The outcomes showed dearth of the lag phase and a considerable change in rate constant, which provides conclusive evidence, that is, heparin-induced BSA aggregation involves the pathway of the downhill polymerization mechanism. Heparin also causes enhancement of fluorescence intensity of BSA significantly. Moreover, heparin was observed to form amyloids and amorphous aggregates of BSA which were confirmed by ThT and ANS fluorescence, respectively. Circular dichroism measurements exhibit a considerable change in the secondary and tertiary structure of the protein due to heparin. In addition, binding studies of heparin with BSA to know the cause of aggregation, isothermal titration calorimetry measurements were exploited, from which heparin was observed to promote the aggregation of BSA by virtue of electrostatic interactions between positively charged amino acid residues of protein and negatively charged groups of GAG. The nature of binding of heparin with BSA is very much apparent with an appreciable heat of interaction and is largely exothermic in nature. Moreover, the Gibbs free energy change (ΔG) is negative, which indicates spontaneous nature of binding, and the enthalpy change (ΔH) and entropy change (ΔS) are also largely negative, which suggest that the interaction is driven by hydrogen bonding.

show abstract

Section: Discussionmentioning

confidence: 99%

Heparin Accelerates the Protein Aggregation via the Downhill Polymerization Mechanism: Multi-Spectroscopic Studies to Delineate the Implications on Proteinopathies

et al. 2021

View full text Add to dashboard Cite

show abstract

“…Biologics have reduced toxic side effects compared to traditional antineoplastic drugs; however, they can overstimulate the immune system, leading to toxicity and inflammation [ 9 , 10 , 11 ]. Additionally, as biologics are mostly composed of proteins and nucleic acids, a change in conformation impairs their function, and they are susceptible to chemical and enzymatic degradation [ 12 , 13 ]. Well-engineered nanocarrier-based DDSs should enhance efficacy and reduce side effects by (1) protecting payloads from degradation and extending their circulation half-lives, (2) enhancing preferential uptake by tumor cells and reduce uptake by non-cancerous cells, and (3) releasing the active payload only when an appropriate stimulus is applied.…”

Section: Introductionmentioning

confidence: 99%

Nanoplatforms for Targeted Stimuli-Responsive Drug Delivery: A Review of Platform Materials and Stimuli-Responsive Release and Targeting Mechanisms

Sun

Davis

2021

Nanomaterials

View full text Add to dashboard Cite

To achieve the promise of stimuli-responsive drug delivery systems for the treatment of cancer, they should (1) avoid premature clearance; (2) accumulate in tumors and undergo endocytosis by cancer cells; and (3) exhibit appropriate stimuli-responsive release of the payload. It is challenging to address all of these requirements simultaneously. However, the numerous proof-of-concept studies addressing one or more of these requirements reported every year have dramatically expanded the toolbox available for the design of drug delivery systems. This review highlights recent advances in the targeting and stimuli-responsiveness of drug delivery systems. It begins with a discussion of nanocarrier types and an overview of the factors influencing nanocarrier biodistribution. On-demand release strategies and their application to each type of nanocarrier are reviewed, including both endogenous and exogenous stimuli. Recent developments in stimuli-responsive targeting strategies are also discussed. The remaining challenges and prospective solutions in the field are discussed throughout the review, which is intended to assist researchers in overcoming interdisciplinary knowledge barriers and increase the speed of development. This review presents a nanocarrier-based drug delivery systems toolbox that enables the application of techniques across platforms and inspires researchers with interdisciplinary information to boost the development of multifunctional therapeutic nanoplatforms for cancer therapy.

show abstract

“…Furthermore, the reversibility of unfolding of these proteins in different formulation conditions might vary 37,40,41 . Recently, it was suggested that one could use ICD experiments to investigate non-reversibility effects during protein unfolding in a denaturant 47,48 . Such experiments study how the apparent Gibbs free energy of protein unfolding changes when different protein concentrations are used to obtain the ICD curves.…”

Section: Introductionmentioning

confidence: 99%

The ReFOLD assay for protein formulation studies and prediction of protein aggregation during long-term storage

Svilenov

Winter

2019

European Journal of Pharmaceutics and Biopharmaceutics

View full text Add to dashboard Cite

The formulation of novel therapeutic proteins is a challenging task which aims at finding formulation conditions that will minimize protein degradation during long-term storage. One particularly important and difficult-to-predict protein degradation pathway is the so-called non-native aggregation. The qualitative and quantitative prediction of the latter has been a subject of extensive research over the past two decades. An increasing body of evidence shows that the widely-used short-term biophysical techniques cannot accurately rank formulation conditions in order of their effect on the aggregation during long-term storage of some therapeutic proteins, e.g. monoclonal antibodies. Here we suggest a novel approach for the selection of formulation conditions that will suppress the formation of protein aggregates during long-term storage. We postulate that conditions (i.e. pH, buffer type, ionic strength) that reduce the isothermal aggregation of various denaturant-induced partially folded protein species will be conditions that impede protein aggregation during long-term storage. To test our hypothesis, we developed an isothermal microdialysis-based unfolding/refolding assay, named ReFOLD, which we use to induce moderate aggregation of partially folded proteins. Next, we assessed the relative monomer yield after isothermal unfolding/refolding of two monoclonal antibodies, each formulated in 12 different conditions. Using the proposed approach, we were able to accurately rank the formulations in order of their effect on the amount of protein aggregates detected after storage for 12 months at 4 ⁰C and 25 ⁰C, while widely-used stability-indicating parameters like protein melting and aggregation onset temperatures failed to provide accurate predictive formulation rankings.

show abstract

Conformational stability and self-association equilibrium in biologics

Cited by 25 publications

References 25 publications

Heparin Accelerates the Protein Aggregation via the Downhill Polymerization Mechanism: Multi-Spectroscopic Studies to Delineate the Implications on Proteinopathies

Heparin Accelerates the Protein Aggregation via the Downhill Polymerization Mechanism: Multi-Spectroscopic Studies to Delineate the Implications on Proteinopathies

Nanoplatforms for Targeted Stimuli-Responsive Drug Delivery: A Review of Platform Materials and Stimuli-Responsive Release and Targeting Mechanisms

The ReFOLD assay for protein formulation studies and prediction of protein aggregation during long-term storage

Contact Info

Product

Resources

About