RBD-Based ELISA and Luminex Predict Anti-SARS-CoV-2 Surrogate-Neutralizing Activity in Two Longitudinal Cohorts of German and Spanish Health Care Workers.

The ability of antibodies to neutralize severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is an important correlate of protection. For routine evaluation of protection, however, a simple and cost-efficient anti-SARS-CoV-2 serological assay predictive of serum neutralizing activity is needed. We analyzed clinical epidemiological data and blood samples from two cohorts of health care workers in Barcelona and Munich to compare several immunological readouts for evaluating antibody levels that could be surrogates of neutralizing activity. We measured IgG levels against SARS-CoV-2 spike protein (S), its S2 subunit, the S1 receptor binding domain (RBD), and the full length and C terminus of nucleocapsid (N) protein by Luminex, and against RBD by enzyme-linked immunosorbent assay (ELISA), and assessed those as predictors of plasma surrogate-neutralizing activity measured by a flow cytometry assay. In addition, we determined the clinical and demographic factors affecting plasma surrogate-neutralizing capacity. Both cohorts showed a high positive correlation between IgG levels to S antigen, especially to RBD, and the levels of plasma surrogate-neutralizing activity, suggesting RBD IgG as a good correlate of plasma neutralizing activity. Symptomatic infection, with symptoms such as loss of taste, dyspnea, rigors, fever and fatigue, was positively associated with anti-RBD IgG positivity by ELISA and Luminex, and with plasma surrogate-neutralizing activity. Our serological assays allow for the prediction of serum neutralization activity without the cost, hazards, time, and expertise needed for surrogate or conventional neutralization assays. Once a cutoff is established, these relatively simple high-throughput antibody assays will provide a fast and cost-effective method of assessing levels of protection from SARS-CoV-2 infection. IMPORTANCE Neutralizing antibody titers are the best correlate of protection against SARS-CoV-2. However, current tests to measure plasma or serum neutralizing activity do not allow high-throughput screening at the population level. Serological tests could be an alternative if they are proved to be good predictors of plasma neutralizing activity. In this study, we analyzed the SARS-CoV-2 serological profiles of two cohorts of health care workers by applying Luminex and ELISA in-house serological assays. Correlations of both serological tests were assessed between them and with a flow cytometry assay to determine plasma surrogate-neutralizing activity. Both assays showed a high positive correlation between IgG levels to S antigens, especially RBD, and the levels of plasma surrogate-neutralizing activity. This result suggests IgG to RBD as a good correlate of plasma surrogate-neutralizing activity and indicates that serology of IgG to RBD could be used to assess levels of protection from SARS-CoV-2 infection.

Predictive Shapes of Ellipsoid PPDL-PTHF Copolymer Particles Prepared by the Phantom Stretching Technique.

Ellipsoidal polymer particles can be prepared from spheres by unidirectional stretching at elevated temperatures, while the particles' aspect ratios (AR) that result from this phantom stretching methodology are often not precisely predictable. Here, an elastic deformation model was exemplarily evaluated for ~50 µm spherical microparticles from PPDL-PTHF block copolymers. The prolate ellipsoidal particles, obtained by stretching in polyvinyl alcohol phantoms, differed in dimensions at identical relative phantoms elongations up to 150%, depending on the relative polymer composition and their systematically altered mechanical properties. Importantly, the resulting particle shapes within the studied range of AR up to ~4 matched the predictions of the elastic deformation model, which includes information of the elastic moduli of phantom and particle materials. These data suggest that the model may be applicable to predict the conditions needed to precisely prepare ellipsoids of desired AR and may be applicable to various deformable particle materials.

In Situ X-Ray Scattering Studies of Poly(ε-caprolactone) Networks with Grafted Poly(ethylene glycol) Chains to Investigate Structural Changes during Dual- and Triple-Shape Effect.

The dual- and triple-shape effects of multiphase polymer networks that contain two crystallizable chain segments have been assessed in situ by combining X-ray measurements with thermomechanical investigations. The studied polymer, named CLEG, is a multiphase polymer network of crystallizable poly(ε-caprolactone) (PCL) with grafted poly(ethylene glycol) (PEG) side chains. Wide-angle (WAXS) and small-angle X-ray scattering (SAXS) measurements were combined with temperature-controlled in situ tensile testing experiments. This integrated approach enables systematic investigation and interpretation of relevant structural features during the programming procedures and the thermally-induced recovery process. Main results concern the combined effect of PCL and PEG crystals on shape fixation, the specific role of low-melting PCL crystallites in the fixation of the low temperature temporary shape, and the different orientation behavior of PCL and PEG crystals during certain stages of the programming procedure. These results demonstrate that crystal orientation effects are dominant for the PCL crystals. The effects of the low temperature PCL crystals could only be investigated when synchrotron radiation was applied. These findings indicate the great potential of in situ X-ray investigations for the creation of design-relevant knowledge about the microscopic foundations of dual- and triple-shape effects in appropriate polymer systems.

Knowledge-based tailoring of gelatin-based materials by functionalization with tyrosine-derived groups.

Molecular models of gelatin-based materials formed the basis for the knowledge-based design of a physically cross-linked polymer system. The computational models with 25 wt.-% water content were validated by comparison of the calculated structural properties with experimental data and were then used as predictive tools to study chain organization, cross-link formation, and estimation of mechanical properties. The introduced tyrosine-derived side groups, desaminotyrosine (DAT) and desaminotyrosyl tyrosine (DATT), led to the reduction of the residual helical conformation and to the formation of physical net-points by π-π interactions and hydrogen bonds. At 25 wt.-% water content, the simulated and experimentally determined mechanical properties were in the same order of magnitude. The degree of swelling in water decreased with increasing the number of inserted aromatic functions, while Young's modulus, elongation at break, and maximum tensile strength increased.

Membrane association and selectivity of the antimicrobial peptide NK-2: a molecular dynamics simulation study.

In an effort to better understand the initial mechanism of selectivity and membrane association of the synthetic antimicrobial peptide NK-2, we have applied molecular dynamics simulation techniques to elucidate the interaction of the peptide with the membrane interfaces. A homogeneous dipalmitoylphosphatidylglycerol (DPPG) and a homogeneous dipalmitoylphosphatidylethanolamine (DPPE) bilayers were taken as model systems for the cytoplasmic bacterial and human erythrocyte membranes, respectively. The results of our simulations on DPPG and DPPE model membranes in the gel phase show that the binding of the peptide, which is considerably stronger for the negatively charged DPPG lipid bilayer than for the zwitterionic DPPE, is mostly governed by electrostatic interactions between negatively charged residues in the membrane and positively charged residues in the peptide. In addition, a characteristic distribution of positively charged residues along the helix facilitates a peptide orientation parallel to the membrane interface. Once the peptides reside close to the membrane surface of DPPG with the more hydrophobic side chains embedded into the membrane interface, the peptide initially disturbs the respective bilayer integrity by a decrease of the order parameter of lipid acyl chain close to the head group region, and by a slightly decrease in bilayer thickness. We found that the peptide retains a high content of helical structure on the zwitterionic membrane-water interface, while the loss of alpha-helicity is observed within a peptide adsorbed onto negatively charged lipid membranes.

Current status of Langmuir monolayer degradation of polymeric biomaterials.

Langmuir monolayer degradation (LMD) experiments with polymers possessing outstanding biomedical application potential yield information regarding the kinetics of their hydrolytic or enzymatic chain scission under well-defined and adjustable degradation conditions. A brief review is given of LMD investigations, including the author's own work on 2-dimensional (2D) polymer systems, providing chain scission data, which are not disturbed by simultaneously occurring transport phenomena, such as water penetration into the sample or transport of scission fragments out of the sample. A knowledge-based approach for the description and simulation of polymer hydrolytic and enzymatic degradation based on a combination of fast LMD experiments and computer simulation of the water penetration is briefly introduced. Finally, the advantages and disadvantages of this approach are discussed.

A molecular dynamic analysis of gelatin as an amorphous material: prediction of mechanical properties of gelatin systems.

Biomaterials are used in regenerative medicine for induced autoregeneration and tissue engineering. This is often challenging, however, due to difficulties in tailoring and controlling the respective material properties. Since functionalization is expected to offer better control, in this study gelatin chains were modified with physically interacting groups based on tyrosine with the aim of causing the formation of physical crosslinks. This method permits application-specific properties like swelling and better tailoring of mechanical properties. The design of the crosslink strategy was supported by molecular dynamic (MD) simulations of amorphous bulk models for gelatin and functionalized gelatins at different water contents (0.8 and 25 wt.-%). The results permitted predictions to be formulated about the expected crosslink density and its influence on equilibrium swelling behavior and on elastic material properties. The models of pure gelatin were used to validate the strategy by comparison between simulated and experimental data such as density, backbone conformation angle distribution, and X-ray scattering spectra. A key result of the simulations was the prediction that increasing the number of aromatic functions attached to the gelatin chain leads to an increase in the number of physical netpoints observed in the simulated bulk packing models. By comparison with the Flory-Rehner model, this suggested reduced equilibrium swelling of the functionalized materials in water, a prediction that was subsequently confirmed by our experimental work. The reduction and control of the equilibrium degree of swelling in water is a key criterion for the applicability of functionalized gelatins when used, for example, as matrices for induced autoregeneration of tissues.

Knowledge-based approach towards hydrolytic degradation of polymer-based biomaterials.

The concept of hydrolytically degradable biomaterials was developed to enable the design of temporary implants that substitute or fulfill a certain function as long as required to support (wound) healing processes or to control the release of drugs. Examples are surgical implants, e.g., sutures, or implantable drug depots for treatment of cancer. In both cases degradability can help to avoid a second surgical procedure for explanation. Although degradable surgical sutures are established in the clinical practice for more than 30 years, still more than 40% of surgical sutures applied in clinics today are nondegradable.1 A major limitation of the established degradable suture materials is the fact that their degradation behavior cannot reliably be predicted by applying existing experimental methodologies. Similar concerns also apply to other degradable implants. Therefore, a knowledge-based approach is clearly needed to overcome the described problems and to enable the tailored design of biodegradable polymer materials. In this Progress Report we describe two methods (as examples for tools for this fundamental approach): molecular modeling combining atomistic bulk interface models with quantum chemical studies and experimental investigations of macromolecule degradation in monolayers on Langmuir-Blodgett (LB) troughs. Finally, an outlook on related future research strategies is provided.

A generalized direct-particle-deletion scheme for the calculation of chemical potential and solubilities of small- and medium-sized molecules in amorphous polymers.

The development, validation, and first applications of a generalized version of an inverse Widom method are described. It permits the calculation of solubility coefficients for molecules as large as, e.g., benzene in all polymers for which reasonable forcefield parameters exist. Predicting the solubility is a key to the knowledge-based design of materials utilized to solve permeability related problems. For long time, particle insertion methods, such as the Widom method, were the only way to predict solubilities from molecular models, but they, in most cases, only worked well for rather small penetrants (e.g., H2, O2, N2). Therefore, a few years ago, a new particle deletion algorithm "DPD" was introduced by Boulougouris, Economou, and Theodorou to overcome this problem in principle. The related computer code was, however, only applicable to special, relatively simple model systems. As application examples for the generalized version described here, solubility calculations for nitrogen, oxygen, and benzene in poly(dimethyl siloxane) are presented.