Characterization of Protein-Protein Interactions via Static and Dynamic Light Scattering

Some, Daniel; Kenrick, Sophia

doi:10.5772/37240

Cited by 22 publications

(33 citation statements)

References 17 publications

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“…Concentration-gradient static light scattering measurements were performed as previously described [19–22] using a Calypso system (Wyatt Technology Corp, Santa Barbara) consisting of a software-controlled multiple syringe pump used to create the concentration gradient, a DAWN-EOS multi-angle light scattering photometer and an Optilab rEX differential refractometer configured to collect data in parallel from the incoming sample stream. The raw data acquired consist of the time-dependent scattering intensity at fourteen scattering angles and the time-dependent differential refractive index.…”

Section: Methodsmentioning

confidence: 99%

“…The recently introduced technique termed composition gradient static light scattering (CG-SLS) has been shown to provide a rapid, high resolution quantitative characterization of macromolecular self- and hetero-association equilibria [19–22]. In the present study we apply this technique to the study of insulin self-association, with two objectives: (1) To check and verify previously reported observations made under a variety of experimental conditions, and (2) to provide additional high resolution data that may be used to test alternative models for equilibrium self-association.…”

Section: Introductionmentioning

confidence: 99%

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pH-dependent self-association of zinc-free insulin characterized by concentration-gradient static light scattering

Attri

Fernández

Minton

2010

Biophysical Chemistry

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Insulin self-association at pH 1.85, 1.95, 3.0, 7.2, 8.0 and 10 was studied via composition gradient light scattering (CG-SLS). At pH 1.95 in acetic acid, insulin was found to exist as a monomer, and in pH 1.85 HCl as a dimer. At pH values of 3.0 -8.0, the dependence of scattering intensity upon total insulin concentration at concentrations of up to 1.5 mg/mL may be quantitatively accounted for by a simple isodesmic association equilibrium scheme requiring only a single association constant for addition of monomer to monomer or any oligomer. At pH 10, the association constant for addition of monomer to monomer was found to be smaller than the association constant for addition of monomer to all higher oligomers by a factor of approximately five. The isodesmic association scheme was also found to quantitatively account for the concentration dependence of the weight-average molecular weight derived from previously published sedimentation equilibrium measurements made at pH 7.0, and the best-fit value of the stepwise equilibrium constant obtained therefrom was in excellent agreement with that obtained from analysis of the light scattering data obtained at pH 7.2

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

pH-dependent self-association of zinc-free insulin characterized by concentration-gradient static light scattering

Attri

Fernández

Minton

2010

Biophysical Chemistry

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“…There are different ways by which B22 value can be measured, including the composition gradient SLS, self interaction chromatography (SIC), osmometry, etc. [144][145][146][147][148][149][150]. Diffusion interaction parameter (Kd) is another useful, dilute solution property that can be utilized for the prediction of high concentration behavior.…”

Section: Analytical Assessment Of High Concentration Formulationmentioning

confidence: 99%

Developments and Challenges for mAb-Based Therapeutics

et al. 2013

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The continuous increase in the number of approved monoclonal antibody (mAb)-based therapy suggests that mAbs, and their derivatives, will continue to be the focus of the biotherapeutics industry for years to come. Although vast improvements in our capability to manufacture, characterize, and stabilize mAbs have been achieved, there are still challenges to be overcome. These include analytical and stabilization approaches associated with the development of high concentration mAb formulations. In addition, several mAb-based modalities are under development, including antibody drug conjugates (ADCs), fusion proteins, and bispecific antibodies (bsAbs), all designed to overcome the limitations encountered with mAb therapy. The current status of their development, with emphasis on manufacturing challenges as well as preliminary clinical results, will be reviewed

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“…Standard biophysical assays such as dynamic light scattering (DLS) and size-exclusion chromatography are limited in their capability in providing simultaneous information regarding conformation and intermolecular interactions in protein solutions (64)(65)(66)(67)(68)(69)(70). Thus, we also employ near-UV circular dichroism (CD) and SANS together to understand how molecular conformation and intermolecular interactions influence solution rheology data, because pH and BSA concentrations are varied.…”

Section: Introductionmentioning

confidence: 99%

Critical Examination of the Colloidal Particle Model of Globular Proteins

Sarangapani¹,

Hudson

Jones

et al. 2015

Biophysical Journal

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Recent studies of globular protein solutions have uniformly adopted a colloidal view of proteins as particles, a perspective that neglects the polymeric primary structure of these biological macromolecules, their intrinsic flexibility, and their ability to sample a large configurational space. While the colloidal perspective often serves as a useful idealization in many cases, the macromolecular identity of proteins must reveal itself under thermodynamic conditions in which the native state is no longer stable, such as denaturing solvents and high protein concentrations where macromolecules tend to have screened excluded volume, charge, and hydrodynamic interactions. Under extreme pH conditions, charge repulsion interactions within the protein chain can overcome the attractive hydrogen-bonding interactions, holding it in its native globular state. Conformational changes can therefore be expected to have great significance on the shear viscosity and other rheological properties of protein solutions. These changes are not envisioned in conventional colloidal protein models and we have initiated an investigation of the scattering and rheological properties of model proteins. We initiate this effort by considering bovine serum albumin because it is a globular protein whose solution properties have also been extensively investigated as a function of pH, temperature, ionic strength, and concentration. As we anticipated, near-ultraviolet circular dichroism measurements and intrinsic viscosity measurements clearly indicate that the bovine serum albumin tertiary structure changes as protein concentration and pH are varied. Our findings point to limited validity of the colloidal protein model and to the need for further consideration and quantification of the effects of conformational changes on protein solution viscosity, protein association, and the phase behavior. Small-angle Neutron Scattering measurements have allowed us to assess how these conformational changes influence protein size, shape, and interprotein interaction strength.

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Characterization of Protein-Protein Interactions via Static and Dynamic Light Scattering

Cited by 22 publications

References 17 publications

pH-dependent self-association of zinc-free insulin characterized by concentration-gradient static light scattering

pH-dependent self-association of zinc-free insulin characterized by concentration-gradient static light scattering

Developments and Challenges for mAb-Based Therapeutics

Critical Examination of the Colloidal Particle Model of Globular Proteins

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