Differential Scanning Calorimetry in the Biopharmaceutical Sciences

Demarest, Stephen J.; Frasca, Verna

doi:10.1016/b978-0-444-59573-7.00011-7

Cited by 4 publications

(3 citation statements)

References 50 publications

(42 reference statements)

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“…We elected to perform differential scanning calorimetry (DSC) to take advantage of the technique's ability to discriminate complex, multi-domain unfolding events (typical for immunoglobulins) due to the derivative nature of the thermal unfolding data and ability to experimentally measure ΔH unfolding . 18 When studied by DSC, both Cκ and Cλ demonstrate single and reversible thermal unfolding curves (albeit 30% reduced ΔH cal for Cλ upon cooling and reheating). The midpoints of thermal denaturation (T m s) were low (58–59 °C) compared to what has generally been observed for intact Fabs.…”

Section: Resultsmentioning

confidence: 99%

Comparing domain interactions within antibody Fabs with kappa and lambda light chains

Toughiri

Ruiz

et al. 2016

mAbs

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IgG antibodies are multi-domain proteins with complex inter-domain interactions. Human IgG heavy chains (HCs) associate with light chains (LCs) of the κ or λ isotype to form mature antibodies capable of binding antigen. The HC/LC interaction involves 4 domains: VH and CH1 from the HC and VL and CL from the LC. Human Fabs with κ LCs have been well characterized for their unfolding behaviors and demonstrate a significant level of cooperativity and stabilization when all 4 domains are intact. Very little is known regarding the thermodynamic properties of human Fabs with λ LCs. Here, we dissect the domain contributions to Fab stability for both κ and λ LC-containing Fabs. We find the cooperativity of unfolding between the constant domains, CH1/Cλ, and variable domains, VH/Vλ, within λ LC-containing Fabs is significantly weaker than that of κ LC-containing Fabs. The data suggests there may not be an evolutionary necessity for strong variable/constant domain cooperativity within λ LC-containing Fabs. After investigating the biophysical properties of Fabs with mismatched variable and constant domain subunits (e.g., VH/Vκ paired with CH1/Cλ or T cell receptor Cα/Cβ), the major role of the constant domains for both κ- and λ-containing Fabs may be to reduce the hydrophobic exposure at the VH/VL interface. Even though Fabs with these non-native pairings were thermodynamically less stable, they secreted well from mammalian cells as well behaved monodisperse proteins, which was in contrast to what was observed with the VH/Vκ and VH/Vλ scFvs that secreted as a mixture of monomer and aggregates.

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Section: Resultsmentioning

confidence: 99%

Comparing domain interactions within antibody Fabs with kappa and lambda light chains

Toughiri

Ruiz

et al. 2016

mAbs

Self Cite

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“…This exercise yielded Δ H vH s of 22–25 kcal/mol for the three reported micelles. The ΔH vH /ΔH cal ratio of one for PEG-P(D,L-Leu)/m-60°C suggests a two-state transition for this micelle [24,49]. On the other hand, we obtained ΔH vH /ΔH cal ratios of 2.5 and 1.7 for the other two types of micelles, which indicates the temperature-induced transition of these micelles is more disorganized.…”

Section: Resultsmentioning

confidence: 83%

Tuning polypeptide-based micellar carrier for efficient combination therapy of ErbB2-positive breast cancer

Soni

Fan

Desale

et al. 2017

Journal of Controlled Release

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“…30,31 DSC analysis DSC is used to measure energy changes during protein denaturation. 32 As shown in Fig. 2(A), the denaturation temperature in T 1 , T 2 , T 3 and T 4 groups was increased compared with the T 0 group.…”

Section: Determination Of the Relative Content Of Mmb And Ombmentioning

confidence: 76%

Effect mechanism of capsaicin and dihydrocapsaicin in chili on the oxidative stability of myoglobin in duck meat

Wu,

Xie,

Zhao

et al. 2024

J Sci Food Agric

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BackgroundMyoglobin (Mb) in duck meat is commenly over‐oxidized when heated at high temperatures, which may worsen the color of the meat. Enhancing the oxidative stability of Mb is essential for improving the color of duck meat. The capsaicin and dihydrocapsaicin (CA‐DI) in Chili exhibited antioxidant properties. Herein, this study investigated effects of CA‐DI on the structure and oxidative damage of Mb by fluorescence spectroscopy, differential scanning calorimetry (DSC) analysis and particle size in duck meat during heat treatment.ResultsWhen the ratio of CA‐DI to Mb was 10: 1 g·kg−1 and heat‐treated for 36 min, oxymyoglobin (OMb) significantly increased, and metmyoglobin (MMb) significantly decreased compared with the control group (P < 0.05). In parallel, the carbonyl content of Mb in CA‐DI group decreased by 43.40 ± 0.10%, the sulfhydryl content increased by 188 ± 0.21%, and the free radical scavenging activity of Mb significantly enhanced (P < 0.05). Moreover, the addition of CA‐DI resulted in a significant decrease in the particle size of the Mb surface (P < 0.05). When the ratio of CA‐DI to Mb was 10: 1 g·kg−1, CA‐DI enhanced the thermal stability and significantly increased thermal denaturation temperature of Mb. The molecular docking results indicated that hydrophobic interactions and hydrogen bonds involved in the binding of CA‐DI to Mb.ConclusionCA‐DI could combine with Mb, improve the oxidation stability of Mb in duck meat. This suggested CA‐DI could be a potential natural antioxidant that improves the color of meat products.This article is protected by copyright. All rights reserved.

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Differential Scanning Calorimetry in the Biopharmaceutical Sciences

Cited by 4 publications

References 50 publications

Comparing domain interactions within antibody Fabs with kappa and lambda light chains

Comparing domain interactions within antibody Fabs with kappa and lambda light chains

Tuning polypeptide-based micellar carrier for efficient combination therapy of ErbB2-positive breast cancer

Effect mechanism of capsaicin and dihydrocapsaicin in chili on the oxidative stability of myoglobin in duck meat

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