Differential Scanning Calorimetry to Quantify Heat-Induced Aggregation in Concentrated Protein Solutions

Jacobs, Matthew; Grace, M. Antony Lilly; Blumlein, Alice; McManus, Jennifer J.

doi:10.1007/978-1-4939-9678-0_9

Cited by 8 publications

(5 citation statements)

References 31 publications

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“…In this particular case, ADA possesses two visible thermal transitions at 74 °C and 84 °C. The main event corresponds to the constant heavy chain-2 and the antigen-binding region unfolding (C H 2 and Fab domains, overlapped), while the second corresponds to the constant heavy chain (C H 3 domain), which is in agreement with the previous literature sources [ 11 , 16 , 17 , 18 ]. This overlapping is due to the close unfolding event of the C H 2 fragment (72–74 °C) with the F ab domain (73–74 °C) [ 16 ].…”

Section: Resultssupporting

confidence: 90%

Stability of Monoclonal Antibodies as Solid Formulation for Auto-Injectors: A Pilot Study

Garcia-Villen,

Gallego,

Sainz-Ramos

et al. 2023

Pharmaceutics

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Drug adherence is a significant medical issue, often responsible for sub-optimal outcomes during the treatment of chronic diseases such as rheumatoid or psoriatic arthritis. Monoclonal antibodies (which are exclusively given parenterally) have been proven to be an effective treatment in these cases. The use of auto-injectors is an effective strategy to improve drug adherence in parenteral treatments since these pen-like devices offer less discomfort and increased user-friendliness over conventional syringe-based delivery. This study aims to investigate the feasibility of including a monoclonal antibody as a solid formulation inside an auto-injector pen. Specifically, the objective was to evaluate the drug stability after a concentration (to reduce the amount of solvent and space needed) and freeze-drying procedure. A preliminary screening of excipients to improve stability was also performed. The nano-DSC results showed that mannitol improved the stability of the concentrated, freeze-dried antibody in comparison to its counterpart without it. However, a small instability of the CH2 domain was still found for mannitol samples, which will warrant further investigation. The present results serve as a stepping stone towards advancing future drug delivery systems that will ultimately improve the patient experience and associated drug adherence.

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Section: Resultssupporting

confidence: 90%

Stability of Monoclonal Antibodies as Solid Formulation for Auto-Injectors: A Pilot Study

Garcia-Villen,

Gallego,

Sainz-Ramos

et al. 2023

Pharmaceutics

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show abstract

“…The differential scanning calorimetry (DSC) peaks of the proteins indicate that different protein structural domains unfold at different temperatures, thus providing information regarding the structural changes during heat‐induced protein unfolding (Jacobs, Grace, Blumlein, & McManus, 2019). Table 2 shows that Peak 1 and Peak 2 obtained via DSC are downward‐absorbing peaks, and when the denaturation temperature of a certain protein is reached, a corresponding absorption peak appears.…”

Section: Resultsmentioning

confidence: 99%

Gel properties of blue round scad (Decapterus maruadsi) mince as influenced by the addition of egg white powder

Wang

et al. 2022

Journal of Texture Studies

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The use of egg white powder (EWP) to enhance the physicochemical properties, molecular structure, and thermal stability of Decapterus maruadsi mince gels was investigated. The thermal stability was analyzed by adding spray‐dried EWP (0, 0.2, 0.4, 0.6, 0.8, and 1%) to the mince, and mince gels were prepared to study the changes in their fracture constant, water distribution, microstructure, and protein conformation of mince gels. In addition, the stress–strain curve of the EWP‐mince gel was measured to obtain its compressive modulus (E). The formation of the mince gel was promoted by EWP, and the whiteness, fracture constant, water‐holding capacity (WHC), and immobilized water were all enhanced. At 0.8% addition of EWP, the fracture constant increased from 176.715 ± 2.463 N/m to 348.631 ± 3.144 N/m (p < .05), which was a nearly twofold improvement. Additionally, the WHC increased from 75.21% to 79.99%, and the percentage of immobilized water increased from 94.03% to 94.91%. The EWP‐mince gel network structure was the most uniform and dense, and there were increases in hydrogen bonds, disulfide bonds, β‐sheets, and β‐turns in mince gels, as well as the storage modulus (G′) and enthalpy (ΔH). In contrast to the control group, the relative content of α‐helixes decreased from 53.34% to 37.09% and transformed into other secondary structures, and the bulk water and cooking loss also decreased to 2.41% and 8.51%, respectively. Consequently, EWP effectively improved the quality of mince products, and the effect was most apparent when 0.8% was added.

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“…PDZ3‐wt, PDZ3‐I389A, and PDZ3‐L349A showed lower reversibility (Fig. S5), which we assumingly assigned to the sample’s prolonged exposure to high temperatures, where the proteins are unfolded and where the exposed hydrophobic residues make the samples aggregation‐prone [36].…”

Section: Resultsmentioning

confidence: 99%

Blocking PSD95‐PDZ3's amyloidogenesis through point mutations that inhibit high‐temperature reversible oligomerization (RO)

et al. 2022

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The third PDZ domain of the postsynaptic density protein 95 (PSD95‐PDZ3; 11 kDa, 103 residues) has a propensity to form amyloid fibrils at high temperatures. At neutral pH, PDZ3 is natively folded, but it exhibits a peculiar three‐state thermal unfolding with a reversible oligomerization (RO) equilibrium at high temperatures, which is uncharacteristic in the unfolding of a small globular protein as PDZ3 is. Here, we examined the RO's role in PDZ3's amyloidogenesis at high‐temperature using two variants (F340A and L342A) that suppress the high‐temperature RO and five single‐alanine‐mutated variants, where we mutated surface‐exposed hydrophobic residues to alanine. Circular Dichroism (CD), Analytical Ultracentrifuge (AUC), and other spectroscopic measurements confirmed the retention of the native structure at ambient temperature. Differential Scanning Calorimetry (DSC) was used to assess the presence or absence of the high‐temperature RO, and the amyloidogenicity of the variants was measured by Thioflavin T (ThT) fluorescence and Transmission Electron Microscopy (TEM). By comparing the fraction of RO and the ThT signal, we found that mutations that suppressed the high‐temperature RO strongly inhibited amyloidogenesis. On the other hand, all variants forming RO also formed amyloids under the same conditions as the wild‐type PDZ3.

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Differential Scanning Calorimetry to Quantify Heat-Induced Aggregation in Concentrated Protein Solutions

Cited by 8 publications

References 31 publications

Stability of Monoclonal Antibodies as Solid Formulation for Auto-Injectors: A Pilot Study

Stability of Monoclonal Antibodies as Solid Formulation for Auto-Injectors: A Pilot Study

Gel properties of blue round scad (Decapterus maruadsi) mince as influenced by the addition of egg white powder

Blocking PSD95‐PDZ3's amyloidogenesis through point mutations that inhibit high‐temperature reversible oligomerization (RO)

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