Survey of chemical unfolding complexity as a unique stability assessment assay for monoclonal antibodies.

Seventy-two intentionally sequence-diverse antibody variable regions were selected, expressed as IgG1 antibodies, and evaluated by chemical unfolding to survey the complexities of denaturant induced unfolding behavior. A two-transition fit well described the curves and uncovered a wide range of sensitivities to denaturant. Four general types of unfolding curves were observed: balanced traces (each transition responsible for half of the total unfolding curve), low-unfolding traces (first transition is a majority of the unfolding curve), high-unfolding traces (the second transition is the majority of the unfolding curve), and coincident traces (the two transitions are found close to each other, approximating a single transition).

AI-based antibody discovery platform identifies novel, diverse, and pharmacologically active therapeutic antibodies against multiple SARS-CoV-2 strains.

Background: We are entering a new era of antibody discovery and optimization where machine learning (ML) processes will become indispensable for the design and development of therapeutics.

Methods: We have constructed a Humanoid Antibody Library for the discovery of therapeutics that is an initial step towards leveraging the utility of artificial intelligence and ML. We describe how we began our validation of the library for antibody discovery by isolating antibodies against a target of pandemic concern, SARS-CoV-2.

Deep Learning in Therapeutic Antibody Development.

Deep learning applied to antibody development is in its adolescence. Low data volumes and biological platform differences make it challenging to develop supervised models that can predict antibody behavior in actual commercial development steps. But successes in modeling general protein behaviors and early antibody models give indications of what is possible for antibodies in general, particularly since antibodies share a common fold.

Evaluation of Crystal Zenith Microtiter Plates for High-Throughput Formulation Screening.

Formulation screening for biotherapeutics can cover a vast array of excipients and stress conditions. These studies consume quantities of limited material and, with higher concentrated therapeutics, more material is needed. Here, we evaluate the use of crystal zenith (CZ) microtiter plates in conjunction with FluoroTec-coated butyl rubber mats as a small-volume, high-throughput system for formulation stability studies.

Framework Mutations of the 10-1074 bnAb Increase Conformational Stability, Manufacturability, and Stability While Preserving Full Neutralization Activity.

The broadly neutralizing anti-HIV antibody, 10-1074, is a highly somatically hypermutated IgG1 being developed for prophylaxis in sub-Saharan Africa. A series of algorithms were applied to identify potentially destabilizing residues in the framework of the Fv region. Of 17 residues defined, a variant was identified encompassing 1 light and 3 heavy chain residues, with significantly increased conformational stability while maintaining full neutralization activity.

Adapting the chemical unfolding assay for high-throughput protein screening using experimental and spectroscopic corrections.

The chemical unfolding (denaturation) assay can be used to calculate the change in the Gibbs free energy of unfolding, ΔG, and inflection point of unfolding, to collectively inform on molecule stability. Here, we evaluated methods for calculating the ΔG across 23 monoclonal antibody sequence variants. These methods are based on how the measured output (intrinsic fluorescence intensity) is treated, including utilizing (a) a single wavelength, (b) a ratio of two wavelengths, (c) a ratio of a single wavelength to an area, and (d) a scatter correction plus a ratio of a single wavelength to an area.

Muscle oxygenation as an indicator of shock severity in patients with suspected severe sepsis or septic shock.

Purpose: The aim of this pilot study was to evaluate the potential of a new noninvasive optical measurement of muscle oxygenation (MOx) to identify shock severity in patients with suspected sepsis.

Methods: We enrolled 51 adult patients in the emergency department (ED) who presented with possible sepsis using traditional Systematic Inflammatory Response Syndrome criteria or who triggered a "Code Sepsis." Noninvasive MOx measurements were made from the first dorsal interosseous muscles of the hand once potential sepsis/septic shock was identified, as soon as possible after admission to the ED.

Muscle oxygenation measurement in humans by noninvasive optical spectroscopy and Locally Weighted Regression.

We have developed a method to make real-time, continuous, noninvasive measurements of muscle oxygenation (Mox) from the surface of the skin. A key development was measurement in both the visible and near infrared (NIR) regions. Measurement of both oxygenated and deoxygenated myoglobin and hemoglobin resulted in a more accurate measurement of Mox than could be achieved with measurement of only the deoxygenated components, as in traditional near-infrared spectroscopy (NIRS).

Estimation of the age of a weathered mixture of volatile organic compounds.

The ability to predict the amount of time that a light petroleum mixture has been weathered could have many applications, such as aiding forensic investigators in determining the cause and intent of a fire. In our study, an evaporation chamber that permits control of airflow and temperature was constructed and used to weather a model nine-component hydrocarbon mixture. The composition of the mixture was monitored over time by gas chromatography and a variety of chemometric models were explored, including partial least squares (PLS), nonlinear PLS (PolyPLS) and locally weighted regression (LWR or loess).

Loopy MSC: a simple way to improve multiplicative scatter correction.

Multiplicative scatter correction (MSC) is a widely used normalization technique. It aims to correct spectra in such a way that they are as close as possible to a reference spectrum, generally the mean of the data set, by changing the scale and the offset of the spectra. When there are other differences in the spectra than just a scale and an offset, the mean spectrum changes after MSC.

Multivariate curve resolution applied to infrared reflection measurements of soil contaminated with an organophosphorus analyte.

Multivariate curve resolution (MCR) is a powerful technique for extracting chemical information from measured spectra of complex mixtures. A modified MCR technique that utilized both measured and second-derivative spectra to account for observed sample-to-sample variability attributable to changes in soil reflectivity was used to estimate the spectrum of dibutyl phosphate (DBP) adsorbed on two different soil types. This algorithm was applied directly to measurements of reflection spectra of soils coated with analyte without resorting to soil preparations such as grinding or dilution in potassium bromide.