A liquid chromatography/mass spectrometry method for screening disulfide tethering fragments

We report the refinement of a high-throughput, liquid chromatography/mass spectrometry (LC/MS)-based screening method for the identification of covalent small-molecule binders to proteins. Using a custom library of 1600 disulfide-capped fragments targeting surface cysteine residues, we optimize sample preparation, chromatography, and ionization conditions to maximize the reliability and flexibility of the approach. Data collection at a rate of 84 s per sample balances speed with reliability for sustained screening over multiple, diverse projects run over a 24-month period. The method is applicable to protein targets of various classes and a range of molecular masses. Data are processed in a custom pipeline that calculates a percent bound value for each compound and identifies false positives by calculating significance of detected masses (signal significance). An example pipeline is available through Biovia's ScienceCloud Protocol Exchange. Data collection and analysis methods for the screening of covalent adducts of intact proteins are now fast enough to screen the largest covalent compound libraries in 1 to 2 days

A liquid chromatography/mass spectrometry method for screening disulfide tethering fragments

We report the refinement of a high-throughput, liquid chromatography/mass spectrometry (LC/MS)-based screening method for the identification of covalent small-molecule binders to proteins. Using a custom library of 1600 disulfide-capped fragments targeting surface cysteine residues, we optimize sample preparation, chromatography, and ionization conditions to maximize the reliability and flexibility of the approach. Data collection at a rate of 84 s per sample balances speed with reliability for sustained screening over multiple, diverse projects run over a 24-month period. The method is applicable to protein targets of various classes and a range of molecular masses. Data are processed in a custom pipeline that calculates a percent bound value for each compound and identifies false positives by calculating significance of detected masses (signal significance). An example pipeline is available through Biovia's ScienceCloud Protocol Exchange. Data collection and analysis methods for the screening of covalent adducts of intact proteins are now fast enough to screen the largest covalent compound libraries in 1 to 2 days

Post-bifurcation analysis of a thin-walled hyperelastic tube under inflation

We consider the problem of bulging, or necking, of an infinite thin-walled hyperelastic tube that is inflated by an internal pressure, with the axial stretch at infinity maintained at unity. We present a simple procedure that can be used to derive the bifurcation condition and to determine the near-critical behaviour analytically. It is shown that there is a bifurcation with zero mode number and that the associated axial variation of near-critical bifurcated configurations is governed by a first-order differential equation that admits a locally bulging or necking solution. This result suggests that the corresponding bifurcation pressure can be identified with the so-called initiation pressure which featured in recent experimental studies. This is supported by good agreement between our theoretical predictions and one set of experimental data. It is also shown that the Gent material model can support both bulging and necking solutions whereas the Varga and Ogden material models can only support bulging solutions. Relevance of the present method to the study of non-linear wave propagation in a fluid-filled distensible tube is also discussed