Multiplexed Antibody Glycosylation Profiling Using Dual Enzyme Digestion and Liquid Chromatography-Triple Quadrupole Mass Spectrometry Method.

Antibody glycosylation plays a crucial role in the humoral immune response by regulating effector functions and influencing the binding affinity to immune cell receptors. Previous studies have focused mainly on the immunoglobulin G (IgG) isotype owing to the analytical challenges associated with other isotypes. Thus, the development of a sensitive and accurate analytical platform is necessary to characterize antibody glycosylation across multiple isotypes. In this study, we have developed an analytical workflow using antibody-light-chain affinity beads to purify IgG, IgA, and IgM from 16 µL of human plasma. Dual enzymes, trypsin and Glu-C, were used during on-bead digestion to obtain enzymatic glycopeptides and protein-specific surrogate peptides. Ultra-high-performance liquid chromatography coupled with triple quadrupole mass spectrometry was used in order to determine the sensitivity and specificity. Our platform targets 95 glycopeptides across the IgG, IgA, and IgM isotypes, as well as eight surrogate peptides representing total IgG, four IgG classes, two IgA classes, and IgM. Four stable isotope-labeled internal standards were added after antibody purification to calibrate the preparation and instrumental bias during analysis. Calibration curves constructed using serially diluted plasma samples showed good curve fitting (R2 > 0.959). The intrabatch and interbatch precision for all the targets had relative standard deviation of less than 29.6%. This method was applied to 19 human plasma samples, and the glycosylation percentages were calculated, which were comparable to those reported in the literature. The developed method is sensitive and accurate for Ig glycosylation profiling. It can be used in clinical investigations, particularly for detailed humoral immune profiling.

Current Research on Probiotics and Fermented Products.

The history of probiotics and fermented products has evolved over millennia [...].

A Review of Plant-Based Drinks Addressing Nutrients, Flavor, and Processing Technologies.

Plant-based drinks have garnered significant attention as viable substitutes for traditional dairy milk, providing options for individuals who are lactose intolerant or allergic to dairy proteins, and those who adhere to vegan or vegetarian diets. In recent years, demand for plant-based drinks has expanded rapidly. Each variety has unique characteristics in terms of flavor, texture, and nutritional composition, offering consumers a diverse range of choices tailored to meet individual preferences and dietary needs. In this review, we aimed to provide a comprehensive overview of the various types of plant-based drinks and explore potential considerations including their nutritional compositions, health benefits, and processing technologies, as well as the challenges facing the plant-based drink processing industry. We delve into scientific evidence supporting the consumption of plant-based drinks, discuss their potential roles in meeting dietary requirements, and address current limitations and concerns regarding their use. We hope to illuminate the growing significance of plant-based drinks as sustainable and nutritious alternatives to dairy milk, and assist individuals in making informed choices regarding their dietary habits, expanding potential applications for plant-based drinks, and providing necessary theoretical and technical support for the development of a plant-based drink processing industry.