The Stability of a Three-State Unfolding Protein

Yang, Binsheng

doi:10.5772/21572

Cited by 1 publication

(2 citation statements)

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“…A multi-state transition model was used to fit each of the triplicate sets of 32 concentrations, giving triplicate fit results for each molecule. The three-state (two transition) equation used to fit the normalized intensity, I norm is based on BinSheng et al [31]. where z i is the fractional contribution of the given transition to the intensity, S f and S u are defined as: and represent the intrinsic intensities of the folded and unfolded states (as y f and y u respectively) and linear correction terms (as m f and m u ) relative to the concentration of denaturant, [D] .…”

Section: Methodsmentioning

confidence: 99%

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Survey of Chemical Unfolding Complexity as a Unique Stability Assessment Assay for Monoclonal Antibodies

Floyd,

Shaver

2024

Preprint

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Seventy-two intentionally sequence-diverse antibody variable regions were selected, expressed as IgG1 antibodies, and evaluated by chemical unfolding to survey the complexities of denaturant induced unfolding behavior. A two-transition fit well described the curves and uncovered a wide range of sensitivities to denaturant. Four general types of unfolding curves were observed: balanced traces (each transition responsible for half of the total unfolding transition), low-unfolding traces (first transition is a majority of the unfolding curve), high-unfolding traces (the second transition is the majority of the unfolding curve), and coincident traces (the two transitions are found close to each other, approximating a single transition).The complexity of the data from this survey indicates that focusing on the first inflection point or fitting a single transition model is likely an over-simplistic method for measuring stability by the chemical unfolding assay. Additionally, other conformational assays, such as thermal and low pH unfolding, showed no correlation with the chemical unfolding results, indicating that each of these assays provide alternate information on the different pathways of antibody conformational stability. These results provide a basis for beginning to better connect unfolding behavior to other physical phenotypic behaviors and production process behaviors.

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