Measurements of Protein-Protein Interactions by Size Exclusion Chromatography

Bloustine, J.; Berejnov, Viatcheslav; Fraden, Seth

doi:10.1016/s0006-3495(03)74684-0

Cited by 58 publications

(80 citation statements)

References 12 publications

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“…In fact, there are quite a few sets of experiments pertinent to our analysis. 14,[20][21][22][23][24][25][26][27][28] It turns out that there is appreciable scatter in the data if we plot all measurements of B 2 at a pH of about 4.5 as a function of ionic strength I ͓NaClϩsmall amount of Na acetate; we have set the ionic strength arising from the latter equal to 0.6ϫconcentration ͑Ref. 21͔͒ ͑see Fig.…”

Section: B Application To Lysozyme 1 Experimental Datamentioning

confidence: 99%

Optimized Baxter model of protein solutions: Electrostatics versus adhesion

Prinsen

Odijk

2004

The Journal of Chemical Physics

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A theory is set up of spherical proteins interacting by screened electrostatics and constant adhesion, in which the effective adhesion parameter is optimized by a variational principle for the free energy. An analytical approach to the second virial coefficient is first outlined by balancing the repulsive electrostatics against part of the bare adhesion. A theory similar in spirit is developed at nonzero concentrations by assuming an appropriate Baxter model as the reference state. The first-order term in a functional expansion of the free energy is set equal to zero which determines the effective adhesion as a function of salt and protein concentrations. The resulting theory is shown to have fairly good predictive power for the ionic-strength dependence of both the second virial coefficient and the osmotic pressure or compressibility of lysozyme up to about 0.2 volume fraction.

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Section: B Application To Lysozyme 1 Experimental Datamentioning

confidence: 99%

Optimized Baxter model of protein solutions: Electrostatics versus adhesion

Prinsen

Odijk

2004

The Journal of Chemical Physics

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“…Of all the analytical techniques used to obtain values for B 22 , such as SLS (George and Wilson 1994), membrane osmometry (Bonnete et al 1999), SE-AUC (Behlke and Ristau 1999), self-interaction chromatography (SIC) (Tessier et al 2002), and SEC-based methods (Bloustine et al 2003), SLS has been the most widely used technique. SLS measures the time-averaged scattering intensity where the excess Rayleigh ratio, R θ , i.e.…”

Section: Second Virial Coefficient (B 22 ): Static Light Scattering (mentioning

confidence: 99%

Assessment and significance of protein–protein interactions during development of protein biopharmaceuticals

Yadav¹,

Li²,

Scherer³

et al. 2013

Biophys Rev

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Early development of protein biotherapeutics using recombinant DNA technology involved progress in the areas of cloning, screening, expression and recovery/purification. As the biotechnology industry matured, resulting in marketed products, a greater emphasis was placed on development of formulations and delivery systems requiring a better understanding of the chemical and physical properties of newly developed protein drugs. Biophysical techniques such as analytical ultracentrifugation, dynamic and static light scattering, and circular dichroism were used to study protein-protein interactions during various stages of development of protein therapeutics. These studies included investigation of protein self-association in many of the early development projects including analysis of highly glycosylated proteins expressed in mammalian CHO cell cultures. Assessment of protein-protein interactions during development of an IgG1 monoclonal antibody that binds to IgE were important in understanding the pharmacokinetics and dosing for this important biotherapeutic used to treat severe allergic IgE-mediated asthma. These studies were extended to the investigation of monoclonal antibodyantigen interactions in human serum using the fluorescent detection system of the analytical ultracentrifuge. Analysis by sedimentation velocity analytical ultracentrifugation was also used to investigate competitive binding to monoclonal antibody targets. Recent development of high concentration protein formulations for subcutaneous administration of therapeutics posed challenges, which resulted in the use of dynamic and static light scattering, and preparative analytical ultracentrifugation to understand the self-association and rheological properties of concentrated monoclonal antibody solutions.

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“…2 Various types of protein aggregates can develop, and a number of analytical methods are required to analyze the entire size range and types -native or unfolded, covalent or noncovalent, reversible or irreversible -of proalso known as A 2 ) is a widely used parameter to study net protein-protein interactions and to describe thermodynamic nonideality of protein solutions. Derived from the virial expansion of the osmotic pressure term, B 22 represents the nonideality in dilute colloid solutions and is given by 7…”

Section: Introductionmentioning

confidence: 99%

“…), which might be repulsive or attractive. 8 Positive B 22 values indicate repulsive protein-protein interactions. In contrast, net attractive protein-protein interactions are given by negative B 22 values, which can reflect a more aggregation-prone system.…”

Section: Introductionmentioning

confidence: 99%