Capillary mediated vitrification is a novel technique that enables storage of antibody critical reagents at ambient temperature: Impact on binding, structure, and laboratory sustainability.

Antibodies and antibody conjugates are essential components of life science research, but their inherent instability necessitates cold storage or lyophilization, posing logistical and sustainability challenges. Capillary-mediated vitrification has shown promise as a tool for improving biomolecule stability. In this study, we assess the feasibility of shipping and storing CMV-stabilized antibody reagents at ambient temperature using a purified rabbit polyclonal as a model system.

Use of capillary-mediated vitrification to produce thermostable, single-use antibody conjugates as immunoassay reagents.

The performance of enzyme-linked immunoassays is directly dependent on the storage, handling, and long-term stability of the critical reagents used in the assay. Currently, antibody reagents are routinely stored as concentrated, multi-use, frozen aliquots. This practice results in material waste, adds complexity to laboratory workflows, and can compromise reagents via cross-contamination and freeze-thaw damage.

Capillary-Mediated Vitrification: Preservation of mRNA at Elevated Temperatures.

RNA is a fundamental tool for molecular and cellular biology research. The recent COVID-19 pandemic has proved it is also invaluable in vaccine development. However, the need for cold storage to maintain RNA integrity and the practical and economic burden associated with cold chain logistics highlight the need for new and improved preservation methods.

Capillary-Mediated Vitrification: A Novel Approach for Improving Thermal Stability of Enzymes and Proteins.

Capillary-mediated vitrification (CMV) is a novel method for stabilizing biological molecules and complexes. CMV leverages capillary evaporation to enable rapid desiccation of aqueous solutions while avoiding both freezing and boiling. In the CMV process, an aqueous solution containing the biological material of interest and common excipients is applied to a solid, porous support, referred to as the scaffold, and desiccated under vacuum.

Macrophages Promote Circulating Tumor Cell-Mediated Local Recurrence following Radiotherapy in Immunosuppressed Patients.

Although radiotherapy (RT) decreases the incidence of locoregional recurrence in breast cancer, patients with triple-negative breast cancer (TNBC) have increased risk of local recurrence following breast-conserving therapy. The relationship between RT and local recurrence is unknown. Here, we tested the hypothesis that recurrence in some instances is due to the attraction of circulating tumor cells to irradiated tissues.

A role of the adaptive immune system in glucose homeostasis.

Objective: The immune system, including the adaptive immune response, has recently been recognized as having a significant role in diet-induced insulin resistance. In this study, we aimed to determine if the adaptive immune system also functions in maintaining physiological glucose homeostasis in the absence of diet-induced disease.

Research Design And Methods: SCID mice and immunocompetent control animals were phenotypically assessed for variations in metabolic parameters and cytokine profiles.

Reactive Oxygen Species Imaging in a Mouse Model of Inflammatory Bowel Disease.

Purpose: Reactive oxygen species (ROS) are important contributors to inflammatory bowel disease (IBD); however, there are insufficient tools for their in vivo evaluation.

Procedures: To determine if a chemiluminescent ROS reporter, coelenterazine, would be a useful tool for the detection of immune cell activation, the macrophage cell line (RAW 264.7) was treated with phorbol myristate acetate (PMA).

Chemiluminescence Imaging of Superoxide Anion Detects Beta-Cell Function and Mass.

Superoxide anion is produced during normal cellular respiration and plays key roles in cellular physiology with its dysregulation being associated with a variety of diseases. Superoxide anion is a short-lived molecule and, therefore, its homeostatic regulation and role in biology and disease requires dynamic quantification with fine temporal resolution. Here we validated coelenterazine as a reporter of intracellular superoxide anion concentration and used it as a dynamic measure both in vitro and in vivo.

Multimodality Imaging of Cancer Superoxide Anion Using the Small Molecule Coelenterazine.

Purpose: We evaluated the small molecule coelenterazine as a potential reporter of cancer-associated superoxide anion in cell culture and in mice.

Procedures: The superoxide anion concentrations of various cancer cell lines were quantified by coelenterazine chemiluminescence in vitro. Coelenteramide fluorescence was detected via flow cytometry and fluorescent microscopy.

Differential fates of biomolecules delivered to target cells via extracellular vesicles.

Extracellular vesicles (EVs), specifically exosomes and microvesicles (MVs), are presumed to play key roles in cell-cell communication via transfer of biomolecules between cells. The biogenesis of these two types of EVs differs as they originate from either the endosomal (exosomes) or plasma (MVs) membranes. To elucidate the primary means through which EVs mediate intercellular communication, we characterized their ability to encapsulate and deliver different types of macromolecules from transiently transfected cells.

Visualizing cellular interactions with a generalized proximity reporter.

Interactions among neighboring cells underpin many physiological processes ranging from early development to immune responses. When these interactions do not function properly, numerous pathologies, including infection and cancer, can result. Molecular imaging technologies, especially optical imaging, are uniquely suited to illuminate complex cellular interactions within the context of living tissues in the body.