Chromatographic measurement of interactions between unlike proteins

Teske, Christopher A.; Blanch, Harvey W.; Prausnitz, John M.

doi:10.1016/j.fluid.2004.01.025

Cited by 26 publications

(37 citation statements)

References 31 publications

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“…27 From a computational perspective, however, lysozyme and chymotrypsinogen are attractive to study for two reasons. First, at 129 and 245 residues respectively, they are both reasonably small, which means that comparatively small simulation boxes can be used in the PMF calculations; given that explicit-solvent CG simulations can still be computationally expensive to perform, this is a significant advantage.…”

Section: Discussionmentioning

confidence: 99%

Toward Optimized Potential Functions for Protein–Protein Interactions in Aqueous Solutions: Osmotic Second Virial Coefficient Calculations Using the MARTINI Coarse-Grained Force Field

Stark

Andrews

Elcock

2013

J. Chem. Theory Comput.

147

210

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Coarse-grained (CG) simulation methods are now widely used to model the structure and dynamics of large biomolecular systems. One important issue for using such methods – especially with regard to using them to model, for example, intracellular environments – is to demonstrate that they can reproduce experimental data on the thermodynamics of protein-protein interactions in aqueous solutions. To examine this issue, we describe here simulations performed using the popular coarse-grained MARTINI force field, aimed at computing the thermodynamics of lysozyme and chymotrypsinogen self-interactions in aqueous solution. Using molecular dynamics simulations to compute potentials of mean force between a pair of protein molecules, we show that the original parameterization of the MARTINI force field is likely to significantly overestimate the strength of protein-protein interactions to the extent that the computed osmotic second virial coefficients are orders of magnitude more negative than experimental estimates. We then show that a simple down-scaling of the van der Waals parameters that describe the interactions between protein pseudo-atoms can bring the simulated thermodynamics into much closer agreement with experiment. Overall, the work shows that it is feasible to test explicit-solvent CG force fields directly against thermodynamic data for proteins in aqueous solutions, and highlights the potential usefulness of osmotic second virial coefficient measurements for fully parameterizing such force fields.

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Section: Discussionmentioning

confidence: 99%

Toward Optimized Potential Functions for Protein–Protein Interactions in Aqueous Solutions: Osmotic Second Virial Coefficient Calculations Using the MARTINI Coarse-Grained Force Field

Stark

Andrews

Elcock

2013

J. Chem. Theory Comput.

147

210

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“…Interestingly, this alternative form of SIC --referred to as cross-interaction chromatography (CIC) [22][23][24] --is based on the premise that crossinteractions between mAbs and polyclonal antibodies are similar to mAb self-interactions. While exceptions to this simple hypothesis are expected, it is interesting that CIC is able to identify mAbs with both low and high propensity to selfassociate, which has been verified via complementary assays [18,22,25].…”

Section: Measurements Of Mab Self-interactionsmentioning

confidence: 99%

Emerging methods for identifying monoclonal antibodies with low propensity to self-associate during the early discovery process

Tessier

Dickinson

2014

Expert Opinion on Drug Delivery

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“…For example, crossinteraction chromatography (CIC) probes non-specific interactions between mAbs and immobilized polyclonal antibodies. [9][10][11][12] Delayed antibody elution indicates a propensity of mAbs to interact non-specifically with polyclonal antibodies, which is correlated with poor mAb solubility. 10,11 A related ELISA method evaluates non-specific interactions between mAbs and immobilized baculovirus particles (BVPs), and such interactions are in turn correlated with fast clearance in vivo.…”

Section: Introductionmentioning

confidence: 99%

An alternative assay to hydrophobic interaction chromatography for high-throughput characterization of monoclonal antibodies

Estep

Caffry

et al. 2015

mAbs

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(2015) An alternative assay to hydrophobic interaction chromatography for high-throughput characterization of monoclonal antibodies, mAbs, 7:3, 553-561, DOI: 10.1080DOI: 10. /19420862.2015 To link to this article: https://doi.org/10. 1080/19420862.2015 Keywords: hydrophobic interaction chromatography, HIC, self-interaction nanoparticle spectroscopy, aggregation, developability, manufacturability, self-association, high-throughput screening, solubility, viscosityThe effectiveness of therapeutic monoclonal antibodies (mAbs) is governed not only by their bioactivity, but also by their biophysical properties. Assays for rapidly evaluating the biophysical properties of mAbs are valuable for identifying those most likely to exhibit superior properties such as high solubility, low viscosity and slow serum clearance. Analytical hydrophobic interaction chromatography (HIC), which is performed at high salt concentrations to enhance hydrophobic interactions, is an attractive assay for identifying mAbs with low hydrophobicity. However, this assay is low throughput and thus not amenable to processing the large numbers of mAbs that are commonly generated during antibody discovery. Therefore, we investigated whether an alternative, higher throughput, assay could be developed that is based on evaluating antibody self-association at high salt concentrations using affinitycapture self-interaction nanoparticle spectroscopy (AC-SINS). Our approach is to coat gold nanoparticles with polyclonal anti-human antibodies, use these conjugates to immobilize human mAbs, and evaluate mAb self-interactions by measuring the plasmon wavelengths of the antibody conjugates as a function of ammonium sulfate concentration. We find that hydrophobic mAbs, as identified by HIC, generally show significant self-association at low to moderate ammonium sulfate concentrations, while hydrophilic mAbs typically show self-association only at high ammonium sulfate concentrations. The correlation between AC-SINS and HIC measurements suggests that our assay, which can evaluate tens to hundreds of mAbs in a parallel manner and requires only small (microgram) amounts of antibody, will enable early identification of mAb candidates with low hydrophobicity and improved biophysical properties.

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Chromatographic measurement of interactions between unlike proteins

Cited by 26 publications

References 31 publications

Toward Optimized Potential Functions for Protein–Protein Interactions in Aqueous Solutions: Osmotic Second Virial Coefficient Calculations Using the MARTINI Coarse-Grained Force Field

Toward Optimized Potential Functions for Protein–Protein Interactions in Aqueous Solutions: Osmotic Second Virial Coefficient Calculations Using the MARTINI Coarse-Grained Force Field

Emerging methods for identifying monoclonal antibodies with low propensity to self-associate during the early discovery process

An alternative assay to hydrophobic interaction chromatography for high-throughput characterization of monoclonal antibodies

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