Methodologies for the rapid determination of the diffusion interaction parameter using Taylor dispersion analysis

Latunde-Dada, Seyi; Bott, Rachel; Barker, D. J.; Leszczyszyn, Oksana I.

doi:10.1039/c5ay02614h

Cited by 14 publications

(10 citation statements)

References 33 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Recently, the use of the interaction parameter k D has gained favor as a more high-throughput means to quantify protein-protein interactions (17). The interaction parameter can be obtained directly with such techniques as dynamic light scattering (DLS) (17,18) or Taylor dispersion analysis (19) by measuring the dependence of protein diffusivity, D m,2 , on protein concentration, c:…”

Section: Introductionmentioning

confidence: 99%

Challenges in Predicting Protein-Protein Interactions from Measurements of Molecular Diffusivity

Sorret

DeWinter

Schwartz

et al. 2016

Biophysical Journal

View full text Add to dashboard Cite

Dynamic light scattering can be used to measure the diffusivity of a protein within a formulation. The dependence of molecular diffusivity on protein concentration (traditionally expressed in terms of the interaction parameter k) is often used to infer whether protein-protein interactions are repulsive or attractive, resulting in solutions that are colloidally stable or unstable, respectively. However, a number of factors unrelated to intermolecular forces can also impact protein diffusion, complicating this interpretation. Here, we investigate the influence of multicomponent diffusion in a ternary protein-salt-water system on protein diffusion and k in the context of Nernst-Planck theory. This analysis demonstrates that large changes in protein diffusivity with protein concentration can result even for hard-sphere systems in the absence of protein-protein interactions. In addition, we show that dynamic light scattering measurements of diffusivity made at low ionic strength cannot be reliably used to detect protein conformational changes. We recommend comparing experimentally determined k values to theoretically predicted excluded-volume contributions, which will allow a more accurate assessment of protein-protein interactions.

show abstract

Section: Introductionmentioning

confidence: 99%

Challenges in Predicting Protein-Protein Interactions from Measurements of Molecular Diffusivity

Sorret

DeWinter

Schwartz

et al. 2016

Biophysical Journal

View full text Add to dashboard Cite

show abstract

“…60,61 As the hydrodynamic diameters observed for (1) MAN with and without Nile red were close to the lower limit of detection for DLS, Taylor dispersion analysis (TDA) of (1) MAN was then carried out to obtain a more accurate estimation of its hydrodynamic size. TDA is an absolute method to determine the diffusion coefficients (D), and thus the hydrodynamic radii (R h ), of molecules in solution, 62,63 based on the dispersion of a solute plug through a uniform cylindrical tube under laminar Poiseuille flow. 64 Following injection, the UV absorbance (λ = 214 nm) of the species in solution is recorded at two detection windows positioned along the capillary (windows 1 and 2, Fig.…”

Section: Resultsmentioning

confidence: 99%

“…64 Importantly for our study, the deconvolution of taylorgrams from mixtures of species in solution allows the estimation of the hydrodynamic radii of their individual components and their relative proportions, 62,67 and has been used to investigate the aggregation of macromolecules in solution. 63,64 For example, Latunde-Dada et al estimated accurately size and relative proportion of the bovine serum albumin (BSA) monomer and thermally-induced aggregates in BSA : (BSA aggregates) in the 3-100% aggregation range. 63 Here, experiments were carried out at a concentration of linear (1) MAN of 2.0 mg mL −1 and at variable [glycopolymer] : [Nile red] molar ratios (Fig.…”

Section: Resultsmentioning

confidence: 99%

Poly(triazolyl methacrylate) glycopolymers as potential targeted unimolecular nanocarriers

et al. 2019

View full text Add to dashboard Cite

show abstract

“…Furthermore, the concentration dependence of mutual diffusion coefficients is widely used to characterize the behavior of molecules in solution and, in particular, to identify conditions where molecular interactions are most favorable in terms of stability. This is because typically, the strengths of these interactions become more pronounced with increasing solute concentration as the solution tends to non-ideality [10], thereby leading to a dependence of the diffusion coefficient on solute concentration. This is of great importance in the development of biopharmaceutical drugs [11][12][13] where it can be used to determine the second virial coefficient (B 2 ), which is a measure of the strength of protein-protein interactions for example.…”

Section: Introductionmentioning

confidence: 99%

“…The latter can be achieved via concentration titration methods [6][7][8][9] or from a single measurement [10]. Mutual diffusion refers to the fluxes of solute and solvent molecules produced by changes or gradients in the concentration of the solution.…”

Section: Introductionmentioning

confidence: 99%

Theoretical Comparisons of the Concentration-Dependent Diffusion Coefficients from Dynamic Light Scattering and Taylor Dispersion Analysis

Latunde-Dada¹

2016

J Anal Bioanal Tech

View full text Add to dashboard Cite

Dynamic Light Scattering and Taylor Dispersion Analysis are two methods employed for the measurement of the diffusion coefficients and interaction parameters of solutes. For self-associating solutes, the interaction parameter provides a measure for the degree of association and hence the stability of the solutes. Due to the characteristics, peculiar to each method, the measured values are weighted averages and are therefore different from the intrinsic or unweighted values. In this paper, using a simple model for self-association, theoretical expressions for the DLS, TDA and intrinsic diffusion coefficients are derived for self-associating solutes. The corresponding interaction parameters are also derived and compared. As expected, at low concentrations, it was found that the DLS gives higher diffusion coefficients than TDA and vice versa at high concentrations. More interestingly, points of inflection were found in the DLS and TDA diffusion coefficient-concentration curves which imply the presence of minima in measured interaction parameters with the TDA minimum occurring at a lower concentration than for DLS. These are absent in the intrinsic curves. Furthermore, it was found that whilst the intrinsic interaction parameter tends to a non-zero value in the limit of low concentrations, the measured interaction parameters vanish. These trends are important for the interpretation of the results obtained from both measurement methods especially when compared to the intrinsic values. With the interaction parameter being increasingly used as a measure of stability, consideration of these expected trends could prove valuable for explaining experimental data measured at low concentrations. Furthermore, they could prove important when comparisons are made between results from the two measurements which are increasingly being used as orthogonal methods for protein analysis.

show abstract

Methodologies for the rapid determination of the diffusion interaction parameter using Taylor dispersion analysis

Abstract: Using Taylor dispersion analysis to determine diffusion coefficients as a function of concentration in a single measurement.

Cited by 14 publications

References 33 publications

Challenges in Predicting Protein-Protein Interactions from Measurements of Molecular Diffusivity

Challenges in Predicting Protein-Protein Interactions from Measurements of Molecular Diffusivity

Poly(triazolyl methacrylate) glycopolymers as potential targeted unimolecular nanocarriers

Theoretical Comparisons of the Concentration-Dependent Diffusion Coefficients from Dynamic Light Scattering and Taylor Dispersion Analysis

Contact Info

Product

Resources

About