Rapid and Quantitative In Vitro Evaluation of SARS-CoV-2 Neutralizing Antibodies and Nanobodies.

Neutralizing monoclonal antibodies and nanobodies have shown promising results as potential therapeutic agents for COVID-19. Identifying such antibodies and nanobodies requires evaluating the neutralization activity of a large number of lead molecules via biological assays, such as the virus neutralization test (VNT). These assays are typically time-consuming and demanding on-lab facilities. Here, we present a rapid and quantitative assay that evaluates the neutralizing efficacy of an antibody or nanobody within 1.5 h, does not require BSL-2 facilities, and consumes only 8 µL of a low concentration (ng/mL) sample for each assay run. We tested the human angiotensin-converting enzyme 2 (ACE2) binding inhibition efficacy of seven antibodies and eight nanobodies and verified that the IC50 values of our assay are comparable with those from SARS-CoV-2 pseudovirus neutralization tests. We also found that our assay could evaluate the neutralizing efficacy against three widespread SARS-CoV-2 variants. We observed increased affinity of these variants for ACE2, including the ß and γ variants. Finally, we demonstrated that our assay enables the rapid identification of an immune-evasive mutation of the SARS-CoV-2 spike protein, utilizing a set of nanobodies with known binding epitopes.

Rapid and quantitative detection of SARS-CoV-2 specific IgG for convalescent serum evaluation.

Convalescent serum with a high abundance of neutralization IgG is a promising therapeutic agent for rescuing COVID-19 patients in the critical stage. Knowing the concentration of SARS-CoV-2 S1-specific IgG is crucial in selecting appropriate convalescent serum donors. Here, we present a portable microfluidic ELISA technology for rapid (15 min), quantitative, and sensitive detection of anti-SARS-CoV-2 S1 IgG in human serum with only 8 µL sample volume. We first identified a humanized monoclonal IgG that has a high binding affinity and a relatively high specificity towards SARS-CoV-2 S1 protein, which can subsequently serve as the calibration standard of anti-SARS-CoV-2 S1 IgG in serological analyses. We then measured the abundance of anti-SARS-CoV-2 S1 IgG in 16 convalescent COVID-19 patients. Due to the availability of the calibration standard and the large dynamic range of our assay, we were able to identify "qualified donors" for convalescent serum therapy with only one fixed dilution factor (200 ×). Finally, we demonstrated that our technology can sensitively detect SARS-CoV-2 antigens (S1 and N proteins) with pg/mL level sensitivities in 40 min. Overall, our technology can greatly facilitate rapid, sensitive, and quantitative analysis of COVID-19 related markers for therapeutic, diagnostic, epidemiologic, and prognostic purposes.

Evidence that melatonin promotes soybean seedlings growth from low-temperature stress by mediating plant mineral elements and genes involved in the antioxidant pathway.

Melatonin (MT) regulates several physiological activities in plants. However, information on how MT regulates soybean growth under low-temperature (LT) stress is lacking. To better understand how MT promotes plant growth and development under LT stress, we designed this study to evaluate the role of MT pretreatment on soybean seedlings exposed to LT stress. Our results showed that LT stress increased oxidative damage by increasing reactive oxygen species (ROS) accumulation, which affected the growth and development of soybean seedlings. However, the application of 5 µmol L-1 MT significantly decreased the oxidative damage by increasing plant mineral element concentrations and the transcript abundance of antioxidant related genes, which enhanced the decrease in ROS accumulation. These results collectively suggest the involvement of MT in improving LT stress tolerance of soybean seedlings by mediating plant mineral elements and the expression of genes involved in the antioxidant pathway.

High Affinity of Chlorin e6 to Immunoglobulin G for Intraoperative Fluorescence Image-Guided Cancer Photodynamic and Checkpoint Blockade Therapy.

Cancer photodynamic therapy (PDT) represents an attractive local treatment in combination with immunotherapy. Successful cancer PDT relies on image guidance to ensure the treatment accuracy. However, existing nanotechnology for co-delivery of photosensitizers and image contrast agents slows the clearance of PDT agents from the body and causes a disparity between the release profiles of the imaging and PDT agents. We have found that the photosensitizer Chlorin e6 (Ce6) is inherently bound to immunoglobulin G (IgG) in a nanomolarity range of affinity. Ce6 and IgG self-assemble to form the nanocomplexes termed Chloringlobulin (Chlorin e6 + immunoglobulin G). Chloringlobulin enhances the Ce6 concentration in the tumor without changing its elimination half-life in blood. Utilizing the immune checkpoint inhibitor antiprogrammed death ligand 1 (PD-L1) (αPD-L1) to prepare αPD-L1 Chloringlobulin, we have demonstrated a combination of Ce6-based red-light fluorescence image-guided surgery, stereotactic PDT, and PD-L1 blockade therapy of mice bearing orthotopic glioma. In mice bearing an orthotopic colon cancer model, we have prepared another Chloringlobulin that allows intraoperative fluorescence image-guided PDT in combination with PD-L1 and cytotoxic T lymphocyte antigen 4 (CTLA-4) dual checkpoint blockade therapy. The Chloringlobulin technology shows great potential for clinical translation of combinatorial intraoperative fluorescence image-guided PDT and checkpoint blockade therapy.

Ultrafast imaging on the formation of periodic ripples on a Si surface with a prefabricated nanogroove induced by a single femtosecond laser pulse.

This paper reports the ultrafast imaging on the formation of periodic surface ripples induced by a single 800 nm, 50 fs laser pulse. The evolution process is observed on a Si surface with a prefabricated nanogroove. The ripples emerge very quickly, only 3 ps after the laser pulse with a fluence of 0.18 J/cm2 irradiating on the surface, and last for several hundreds of picoseconds. The ultrafast dynamics of laser-matter interaction, such as free carrier excitation, carrier and lattice heating, surface plasmon polariton (SPP) excitation, etc, are studied theoretically. The theoretical and experimental results support that the periodic ripples are caused by the periodic energy deposition due to SPP excitation. The emerge time could identify the surface melting causing the formation of periodic ripples, and exclude the other thermal effects, for example, hydrodynamics.

Coherent beam combination of multiple beams based on near-field angle modulation.

We proposed a new phase-locking technique for multibeam coherent beam combination. By near-field angle modulation and angular spectrum measurement, we obtained the relative phase between each pair of beams with one camera. This method is appropriate for multibeam schemes and possesses the advantages of high accuracy, resistance to energy fluctuation, and simplicity, as shown by the analysis in this study. In a proof-of-principle experiment, we realized the phase-locking of three beams, achieving a Strehl ratio of 89.5%. Our method may supply a scheme for multibeam coherent combining of ultra-intense bulk laser systems.

Multi-region phase calibration of liquid crystal SLM for holographic display.

The liquid crystal spatial light modulator (SLM) is able to provide flexible wavefront control, whereas the initial phase and its response distortions will heavily influence the modulation accuracy. The currently existing calibration methods are tedious and time consuming. A novel multi-region calibration method for minimizing those distortions is proposed. The entire panel is divided into several local regions based on the similarity of phase response characteristic. The nonlinear phase response and static phase distortion of each local region are calibrated in the iterative division procedure. The calibration method is theoretically analyzed and experimentally verified. For the Jasper 4 K SLM panel, when five local regions are built, the root mean error of linear phase shifts is reduced to 0.1 rad and the compensation accuracy of the static phase distortion reaches 0.24 wavelength. The calibrated SLM is applied for the color holographic display and the results show that the reconstructed image quality is improved significantly. The proposed method is simpler and faster because of the reasonable regional division and lower calibration complexity. It could be used for the calibration of various phase only or complex modulators with high space bandwidth product in the future.

Optimizing Nonlinear Optical Visibility of Two-Dimensional Materials.

Two-dimensional (2D) materials have attracted broad research interests across various nonlinear optical (NLO) studies, including nonlinear photoluminescence (NPL), second harmonic generation (SHG), transient absorption (TA), and so forth. These studies have unveiled important features and information of 2D materials, such as in grain boundaries, defects, and crystal orientations. However, as most research studies focused on the intrinsic NLO processes, little attention has been paid to the substrates underneath. Here, we discovered that the NLO signal depends significantly on the thickness of SiO2 in SiO2/Si substrates. A 40-fold enhancement of the NPL signal of graphene was observed when the SiO2 thickness was varied from 270 to 125 nm under 800 nm excitation. We systematically studied the NPL intensity of graphene on three different SiO2 thicknesses within a pump wavelength range of 800-1100 nm. The results agreed with a numerical model based on back reflection and interference. Furthermore, we have extended our measurements to include TA and SHG of graphene and MoS2, confirming that SiO2 thickness has similar effects on all of the three major types of NLO signals. Our results will serve as an important guidance for choosing the optimum substrates to conduct NLO research studies on 2D materials.