SWATH-MS insights on sodium butyrate effect on mAbs production and redox homeostasis in CHO cells.

Sodium butyrate (NaBu), well-known as a histone deacetylase inhibitor and for its capacity to impede cell growth, can enhance the production of a specific protein, such as an antibody, in recombinant Chinese hamster ovary (CHO) cell cultures. In this study, two CHO cell lines, namely K1 and DG44, along with their corresponding mAb-producing lines, K1-Pr and DG44-Pr, were cultivated with or without NaBu. A SWATH-based profiling method was employed to analyze the proteome.

A Novel Crosslinking Approach for Biomanufacturing of a Collagen-Based Skin Dermal Template.

Third-degree burns result in extensive damage to the skin's epidermal and dermal layers, with limited treatment options available. Currently, xenogeneic collagen-based skin grafts are used as scaffolds to integrate into the wound bed and provide a template for neodermis formation. Existing commercial products like Integra dermal templates rely on a time-consuming and variable dehydrothermal (DHT) crosslinking process.

Exploring cost reduction strategies for serum free media development.

Cultivated meat production offers solutions in addressing global food security and sustainability challenges. However, serum-free media (SFM) used in cultivating the cells are expensive, contributing to at least 50% of variable operating costs. This review explores technologies for cost-effective SFM, focusing on reducing cost from using growth factors and recombinant proteins, using affordable raw materials for basal media, and implementing cost-saving measures like media recycling and reducing waste build-up.

Perforated imprinting on high moisture meat analogue confers long range mechanical anisotropy resembling meat cuts.

Meat cuts, when cooked and masticated, separate into fibrous structures because of the long-range mechanical anisotropy (LMA) exhibited by muscle fascicles, which is not fully recapitulated in alternative proteins produced using molecular alignment technology like high moisture extrusion. We have developed a scalable perforated micro-imprinting technology to greatly enhance LMA in high moisture meat analogue (HMMA). By imprinting 1 mm thick HMMA sheets with perforated patterns (optimized by AI), we observed up to 5 × more anisotropic separation of fibrous structures in a one-dimensional pulling LMA analysis, to match the fibrousness of the cooked chicken breast, duck breast, pork loin and beef loin.

Sdd3 regulates the biofilm formation of via the Rho1-PKC-MAPK pathway.

, the most frequently isolated fungal pathogen in humans, forms biofilms that enhance resistance to antifungal drugs and host immunity, leading to frequent treatment failure. Understanding the molecular mechanisms governing biofilm formation is crucial for developing anti-biofilm therapies. In this study, we conducted a genetic screen to identify novel genes that regulate biofilm formation in .

High Extracellular K Skews T-Cell Differentiation Towards Tumour Promoting Th2 and T Subsets.

Potassium ions (K) released from dying necrotic tumour cells accumulate in the tumour microenvironment (TME) and increase the local K concentration to 50 mM (high-[K]). Here, we demonstrate that high-[K] decreases expression of the T-cell receptor subunits CD3ε and CD3ζ and co-stimulatory receptor CD28 and thereby dysregulates intracellular signal transduction cascades. High-[K] also alters the metabolic profiles of T-cells, limiting the metabolism of glucose and glutamine, consistent with functional exhaustion.

Removing immunogenic double-stranded RNA impurities post in vitro transcription synthesis for mRNA therapeutics production: A review of chromatography strategies.

Messenger RNA (mRNA) vaccines and therapeutics hold immense potential for a wide range of clinical applications. However, the in vitro transcription (IVT) process used to synthesize mRNA also results in the generation of a by-product, double-stranded RNA (dsRNA), which can trigger innate immune activation and reduce translation activity. Although various efforts have been made to optimize IVT synthesis to minimize dsRNA formation, dsRNA impurities still cannot be fully resolved.

Device for automated aseptic sampling: Automated sampling solution for future cell and gene manufacturing.

Cell sampling is a key step performed regularly throughout the cell manufacturing process to gather cell samples for cell growth, progress, and characteristics analysis. While the current method of sampling by pipetting in a biosafety cabinet is commonly used, it is labour-intensive and susceptible to contamination risks. We have developed Device for Automated Aseptic Sampling (DAAS), to enable automated, small volume (0.

Cellulose functionalized magnetic beads for high throughput glycosylation analysis in biotherapeutic modalities.

The increasing demand for biotherapeutics has necessitated the evaluation of their critical quality attributes, one of which is glycosylation, an essential post-translational modification found on many biological molecules. In particular, the purification of N-glycans after their release from the proteins and derivatization is important in ensuring the removal of the deglycosylated protein, excess labelling reagents and salts for subsequent analysis. However, current methods of N-glycans purification are either expensive, laborious, time-consuming or not catered for high throughput analysis.

Exome sequencing in Asian populations identifies low-frequency and rare coding variation influencing Parkinson's disease risk.

Parkinson's disease (PD) is an incurable, progressive and common movement disorder that is increasing in incidence globally because of population aging. We hypothesized that the landscape of rare, protein-altering variants could provide further insights into disease pathogenesis. Here we performed whole-exome sequencing followed by gene-based tests on 4,298 PD cases and 5,512 controls of Asian ancestry.

Real-Time Adaptive Inline Acidification Enhances Continuous pH Control for Viral Inactivation.

Existing low pH viral inactivation methods for continuous downstream processing of biologics typically rely on predictive models to estimate the necessary pH adjustments. However, these methods are of limited use during the process development stage due to the dynamic nature of capture chromatography, where batch variations can alter the eluted protein titer. This study introduces an inline viral inactivation system (IVIS) that utilizes real-time adaptive control and inline sensor readings to precisely regulate the pH manipulation for inline acidification and continuous viral inactivation.

Enriching central memory T cells using novel bioreactor design for T cell manufacturing.

Background: The manufacturing of T cell therapies aims to achieve high yields of product with potent phenotypes. We have developed a novel bioreactor, bioreactor with expandable culture area-dual chamber (BECA-D), which has previously demonstrated functionality for scaled T cell manufacturing.

Methods And Results: Methods and Results: In this study, incorporation of a stirring mechanism into the double-chamber bioreactor design was tested to homogenize the media components between the two chambers.

Thermal and pH stress dictate distinct mechanisms of monoclonal antibody aggregation.

Protein aggregation is a significant challenge in the development of monoclonal antibodies (mAbs), which can be exacerbated by stress conditions encountered along its production pipeline. In this study, we examine how thermal and pH stress conditions influence mAb aggregation mechanisms. We observe a complex interplay between these factors that significantly affects mAb stability, particularly under combined stress conditions.

A multiantigenic antibacterial nanovaccine utilizing hybrid membrane vesicles for combating Pseudomonas aeruginosa infections.

Bacterial infections, especially those caused by multidrug-resistant pathogens, pose a significant threat to public health. Vaccines are a crucial tool in fighting these infections; however, no clinically available vaccine exists for the most common bacterial infections, such as those caused by Pseudomonas aeruginosa. Herein, a multiantigenic antibacterial nanovaccine (AuNP@HMV@SPs) is reported to combat P.

Advancements and regulations of biomanufacturing cell-based cartilage repair therapies.

Cell-based therapies for cartilage repair, including autologous chondrocyte implantation and allogeneic stem cell treatments, show great promise but face challenges due to high costs and regulatory hurdles. This review summarizes available and investigational products, focusing on allogeneic therapies and the impact of diverse regulatory landscapes on their clinical translation.

Rapid manufacturing of CAR-T therapy: strategies and impact.

The accessibility of autologous chimeric antigen receptor T cell (CAR-T) therapies is challenged by the complex processes and capacity constraints of manufacturing. Rapid manufacturing capable of shortening manufacturing timelines could transform the CAR-T field. Here, we outline approaches to rapid CAR-T manufacturing, highlighting its impact on various stakeholders in the landscape.

Integrative approach to assessing bioactivity from hempseed protein isolate extracted and dehydrated by different methods: Synergising in silico prediction and in vitro validation.

This study demonstrated a comprehensive workflow combining in silico screening and prediction with in vitro validation to investigate the bioactivity of hempseed protein isolate (HPI) extracted and dehydrated using different methods. By adopting an in silico approach, 13 major proteins of HPI were hydrolysed by 20 selected enzymes, leading to the prediction of 20 potential bioactivities. With papain hydrolysis, dipeptidyl peptidase-IV (DPP4) and angiotensin-converting enzyme (ACE) inhibitory activities emerged as having the highest potential.

Generating and characterizing a comprehensive panel of CHO cells glycosylation mutants for advancing glycobiology and biotechnology research.

This report describes the development and characterization of a comprehensive collection of CHO cell glycosylation mutants with significant potential for advancing glycobiology and biotechnology. EPO-Fc and trastuzumab, two model molecules, were produced using these mutants to assess the effects of mutated glycogenes, and LC-MS/MS analysis was employed to quantitatively analyse their N-glycans. EPO-Fc exhibited exclusively homogeneous Man9 glycans only when nearly all α-mannosidases in the genome were inactivated, except lysosomal MAN2B1.

Dietary amino acids, macronutrients, vaginal birth, and breastfeeding are associated with the vaginal microbiome in early pregnancy.

Unlabelled: The vaginal microbiome is a key player in the etiology of spontaneous preterm birth. This study aimed to illustrate maternal environmental factors associated with vaginal microbiota composition and function in pregnancy. Women in healthy pregnancy had vaginal microbial sampling from the posterior vaginal fornix performed at 16 weeks gestation.

p53-dependent crosstalk between DNA replication integrity and redox metabolism mediated through a NRF2-PARP1 axis.

Mechanisms underlying p53-mediated protection of the replicating genome remain elusive, despite the quintessential role of p53 in maintaining genomic stability. Here, we uncover an unexpected function of p53 in curbing replication stress by limiting PARP1 activity and preventing the unscheduled degradation of deprotected stalled forks. We searched for p53-dependent factors and elucidated RRM2B as a prime factor.

Hybrid modeling for in silico optimization of a dynamic perfusion cell culture process.

The bio-pharmaceutical industry heavily relies on mammalian cells for the production of bio-therapeutic proteins. The complexity of implementing and high cost-of-goods of these processes are currently limiting more widespread patient access. This is driving efforts to enhance cell culture productivity and cost reduction.

Investigation on environmental factors contributing to bispecific antibody stability and the reversal of self-associated aggregates.

Bispecific antibodies (bsAbs) hold promises for enhanced therapeutic potential surpassing that of their parental monoclonal antibodies. However, bsAbs pose great challenges in their manufacturing, and one of the common reasons is their susceptibility to aggregation. Building on previous studies demonstrating the functionality and potential manufacturability of Fab-scFv format bsAb, this investigation delved into the impact of environmental factors-such as pH, buffer types, ionic strength, protein concentrations, and temperatures-on its stability and the reversal of its self-associated aggregates.

Ex vivo T cell differentiation in adoptive immunotherapy manufacturing: Critical process parameters and analytical technologies.

Adoptive immunotherapy shows great promise as a treatment for cancer and other diseases. Recent evidence suggests that the therapeutic efficacy of these cell-based therapies can be enhanced by the enrichment of less-differentiated T cell subpopulations in the therapeutic product, giving rise to a need for advanced manufacturing technologies capable of enriching these subpopulations through regulation of T cell differentiation. Studies have shown that modifying certain critical process control parameters, such as cytokines, metabolites, amino acids, and culture environment, can effectively manipulate T cell differentiation in ex vivo cultures.