Characterizing Reversible Protein Association at Moderately High Concentration Via Composition-Gradient Static Light Scattering

Some, Daniel; Pollastrini, Joseph; Cao, Shawn

doi:10.1016/j.xphs.2016.05.018

Cited by 8 publications

(6 citation statements)

References 15 publications

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“…Static light scattering (SLS) is commonly used to measure protein interactions at dilute conditions via the osmotic second virial coefficient B 22 , and also at high concentration (up to 200 mg/mL) ,− at low wave vector ( q ) values, typically ∼0.0018 Å –1 . The small volume requirements (∼20 μL) and potential for high-throughput measurements using commercial instruments − are relevant for guiding mAb discovery and formulation. SLS provides a quantitative measure of the net interaction strength via the protein–protein Kirkwood–Buff integral G 22 ,,, or zero- q static structure factor S ( q → 0). , As shown in Section 5 of the Supporting Information in eqs S5a–S8, S ( q → 0) is a calculated quantity: .…”

Section: Introductionmentioning

confidence: 99%

Protein–Protein Interactions of Highly Concentrated Monoclonal Antibody Solutions via Static Light Scattering and Influence on the Viscosity

et al. 2019

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The ability to design and formulate mAbs to minimize attractive interactions at high concentrations is important for protein processing, stability, and administration, particularly in subcutaneous delivery, where high viscosities are often challenging. The strength of protein−protein interactions (PPIs) of an IgG1 and IgG4 monoclonal antibody (mAb) from low to high concentration was determined by static light scattering (SLS) and used to understand viscosity data. The PPI were tuned using NaCl and five organic ionic co-solutes. The PPI strength was quantified by the normalized structure factor S(0)/S(0) HS and Kirkwood−Buff integral G 22 /G 22,HS (HS = hard sphere) determined from the SLS data and also by fits with (1) a spherical Yukawa potential and (2) an interacting hard sphere (IHS) model, which describes attraction in terms of hypothetical oligomers. The IHS model was better able to capture the scattering behavior of the more strongly interacting systems (mAb and/or co-solute) than the spherical Yukawa potential. For each descriptor of PPI, linear correlations were obtained between the viscosity at high concentration (200 mg/mL) and the interaction strengths evaluated both at low (20 mg/mL) and high concentrations (200 mg/mL) for a given mAb. However, the only parameter that provided a correlation across both mAbs was the oligomer mass ratio (m oligomer /m monomer+dimer ) from the IHS model, indicating the importance of self-association (in addition to the direct influence of the attractive PPI) on the viscosity.

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Section: Introductionmentioning

confidence: 99%

Protein–Protein Interactions of Highly Concentrated Monoclonal Antibody Solutions via Static Light Scattering and Influence on the Viscosity

et al. 2019

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“…One dilution experiment was performed for each NaCl concentration. Instrument control, data acquisition and data analysis were all carried out with the Calypso software, which implements the protein interaction model fitting, including a single-species effective hard-sphere volume approximation (EHSVA) based on Equations (7)–(10) [ 69 , 70 ]:

where w tot represents the combined mass/volume concentration of the monomers and all oligomers, v is the specific volume, c tot is the total molar concentration of free and bound protein monomers in solution; and

and

are the partial concentrations of free monomers and oligomers, respectively. K i is the equilibrium association constant for the monomer-oligomer association.…”

Section: Methodsmentioning

confidence: 99%

Effects of Monovalent Salt on Protein-Protein Interactions of Dilute and Concentrated Monoclonal Antibody Formulations

Clark

Pollastrini

et al. 2022

Antibodies

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In this study, we used sodium chloride (NaCl) to extensively modulate non-specific protein-protein interactions (PPI) of a humanized anti-streptavidin monoclonal antibody class 2 molecule (ASA-IgG2). The changes in PPI with varying NaCl (CNaCl) and monoclonal antibody (mAb) concentration (CmAb) were assessed using the diffusion interaction parameter kD and second virial coefficient B22 measured from solutions with low to moderate CmAb. The effective structure factor S(q)eff measured from concentrated mAb solutions using small-angle X-ray and neutron scattering (SAXS/SANS) was also used to characterize the PPI. Our results found that the nature of net PPI changed not only with CNaCl, but also with increasing CmAb. As a result, parameters measured from dilute and concentrated mAb samples could lead to different predictions on the stability of mAb formulations. We also compared experimentally determined viscosity results with those predicted from interaction parameters, including kD and S(q)eff. The lack of a clear correlation between interaction parameters and measured viscosity values indicates that the relationship between viscosity and PPI is concentration-dependent. Collectively, the behavior of flexible mAb molecules in concentrated solutions may not be correctly predicted using models where proteins are considered to be uniform colloid particles defined by parameters derived from low CmAb.

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“…118 Among recent applications are studies focused on guiding mAb discovery and formulation by quantifying self-association binding affinities and oligomerization. [119][120][121][122][123] Pallesen et al used CG-MALS to complement BLI studies of Ebola virus GP and antibodies. 124 In 2019, Hastie et al applied CG-MALS to confirm binding affinities of Lassa virus surface glycoprotein (GPC) and antibodies.…”

Section: Composition-gradient Multi-angle Light Scattering (Cg-mals)mentioning

confidence: 99%

Label-free methods for opticalin vitrocharacterization of protein–protein interactions

Soltermann

Struwe

Kukura

2021

Phys. Chem. Chem. Phys.

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Characterizing Reversible Protein Association at Moderately High Concentration Via Composition-Gradient Static Light Scattering

Cited by 8 publications

References 15 publications

Protein–Protein Interactions of Highly Concentrated Monoclonal Antibody Solutions via Static Light Scattering and Influence on the Viscosity

Protein–Protein Interactions of Highly Concentrated Monoclonal Antibody Solutions via Static Light Scattering and Influence on the Viscosity

Effects of Monovalent Salt on Protein-Protein Interactions of Dilute and Concentrated Monoclonal Antibody Formulations

Label-free methods for opticalin vitrocharacterization of protein–protein interactions

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