Biotechnology and Bioengineering最新文献

Meningiomas account for about 40% of all primary brain tumors. How ever effective treatments for recurrent or inoperable cases remain limited. We previously demonstrated that culturing cancer cells on specific hydrogels efficiently induces cancer stem cells across multiple cancer types, a process we termed hydrogel activated reprogramming (HARP) phenomenon. In this study, we aimed to identify key molecules involved in the induction of meningioma stem cells through hydrogel-based culture. Meningioma cells cultured on hydrogels were analyzed for expression of established stem cell markers and for tumorigenicity. Microarray analysis was performed to identify meningioma stem cell specific markers and to evaluate the application of these marker molecules as a therapeutic targets or as diagnostic tools for pathological grading. Canonical stem cell markers including Nanog, and Oct3/4 were upregulated in culturing meningioma cells on hydrogels. Comprehensive gene expression analysis identified some molecules involved in cancer stem cell activity among which CXCR4 was selected as a potential therapeutic target. Stimulation of CXCR4 with its ligand CXCL12 resulted in increased expression of stem cell markers. In human meningioma pathological specimens and cultured cell lines, there was a correlation between CXCR4 expression levels and NF2 mutations and/or deletions. CXCR4 immunohistochemistry was frequently positive in cases with brain invasion along with brain invasion area. These findings suggest that CXCR4 immunohistochemistry may be useful in suggesting typical CNS WHO grade 1 meningiomas without the need for molecular analysis. We have defined meningioma stem cell signature via HARP phenomenon and identified CXCR4 with biological significance as being diagnostic target. IMPORTANCE OF THE STUDY: In addition to morphological evaluation, immunohistochemistry and genetic alteration increasingly incorporated into the diagnostic criteria for central nervous system (CNS) tumors. From the CNS WHO 5th edition onwards, epigenetic features including DNA methylation profiling, have also been adopted as diagnostic criteria. In this study, we induced epigenetic changes in meningioma cells and successfully promoted cancer stemness highlighting the potential importance of this approach for both meningioma research and meningioma diagnostic development. Furthermore, microarray analysis identified CXCR4 as a molecule consistently upregulated during stem cell induction across all three hydrogel conditions. Subsequent analysis revealed that CXCR4 immunohistochemistry may reflect the distribution of meningioma stem cells, supporting its potential utility as a diagnostic marker. By integrating basic experimental findings with histopathological evaluation of clinical specimens, this report will contribute to the advancement of meningioma research and diagnostic strategies.

Coronaviruses within the Merbecovirus subgenus, including Middle East respiratory syndrome coronavirus (MERS-CoV) and its dipeptidyl peptidase 4 (DPP4)-using relatives, pose a persistent zoonotic threat. Efforts to prepare for future Merbecovirus spillover events require vaccines that protect beyond a single virus strain. To evaluate antigenic conservation and cross-protective potential, SpyCatcher-mi3 nanoparticles displaying the receptor-binding domain and subdomain 1 (RBD-SD1) from three DPP4-using merbecoviruses, MERS-CoV, NL140422, and HKU4, were generated. Female mice immunized with these nanoparticle vaccines elicited robust IgG antibody endpoint binding titers and cross-reactive antibody responses against the three merbecoviruses. Only the MERS-CoV RBD-SD1 vaccine, however, elicited neutralizing antibodies against MERS-CoV. While vaccination with MERS-CoV RBD-SD1 reduced lung viral titers in MERS-CoV-challenged human DPP4 mice below the limit of detection, no significant reduction in virus titers was seen in NL140422- and HKU4-RBD-SD1-vaccine-immunized mice. These findings indicate that while the RBD-SD1 interface presents conserved antigenic features sufficient for serological cross-recognition, these epitopes may not be functionally immunodominant for cross-neutralization.

Multimodal chromatography has emerged as a powerful tool for the purification of monoclonal antibodies (mAbs) and their derivatives-including antibody fragments (Fabs), Fc-fusions, bispecific (BsAb), and antibody-drug conjugates (ADCs)-offering enhanced selectivity through the integration of ionic, hydrophobic, hydrogen-bonding, and π-π interactions. This review presents the first comprehensive comparison of all commercially available multimodal chromatography resins used in the purification of antibody-based (Ab-based) products, incorporating experimental data from peer-reviewed literature, supplier documentation, and technical reports. Beyond performance metrics such as binding capacity, recovery, host cell proteins (HCPs) and host cell DNA (hcDNA) clearance, this work synthesizes molecular-level insights into antibody-ligand interactions derived from NMR, DEPC labeling, molecular docking, and thermodynamic analyses. It also compiles data on resin orthogonality, aggregation removal, and the impact of mobile phase modifiers. Recent advances in quantitative structure-property relationship (QSPR) modeling, in silico partition coefficient prediction, and high-throughput screening are discussed as enablers of rational resin selection. This review presents a strategic framework that integrates molecular descriptors, mechanistic understanding, and empirical data to guide the selection and optimization of multimodal chromatography resins, positioning them as essential tools in next-generation biopharmaceutical purification platforms.

Biopharmaceutical manufacturing requires robust analytics and process controls throughout production to insure high yield of quality products. New methodologies for rapidly accessing and integrating data-rich information from complex dynamic biological environments are of great interest. We suggest electronic detection of biological redox signatures based on mediated electrochemical probing (MEP) as an innovative, simple, and rapid modality for interrogating these complex systems. We have previously shown that by leveraging the redox properties of interchain-disulfide bonds within antibodies one can interrogate antibody structure, in particular, the occurrence of partially reduced forms that can occur during production processes. In this work, we expanded on this method to decrease sample-to-answer time and increase detection reliability by optimizing electrical measurements and implementing a machine learning pipeline that intakes the electrochemical data and quantifies free cysteine concentrations as well as reduced antibody fragment levels in growth media. In doing so, we demonstrate a simple method-development platform for electrochemical dataset generation, feature selection, and model optimization that may be transferable to other biological production processes. Further, the developed method offers opportunities for at line digital integration for monitoring complex product attributes throughout bioprocessing.