Biotechnology Journal最新文献

From infectious diseases and cancers to various rare diseases, mRNAs have demonstrated considerable therapeutic potential for a wide range of diseases. However, due to their single-stranded structure, mRNA molecules are vulnerable to enzyme-mediated degradation. Therefore, the inherent instability of mRNA poses a significant challenge. In this review, we explore strategies to slow down the degradation rate, such as removing degradative enzymes, adding protective substances, and optimizing storage and transport conditions to enhance mRNA stability. Furthermore, optimizing the sequence and structure of mRNAs is crucial for improving stability, which can be significantly aided by fine-tuning the sequences of the 5' untranslated region, open reading frame, and 3' untranslated region, along with introducing various RNA modifications. The design of novel mRNA structures, including circular mRNA and self-amplifying RNA, also offers novel approaches for enhancing mRNA stability. Additionally, we briefly introduce the use of mRNA delivery materials for improving stability and discuss current challenges and future directions in mRNA development. With ongoing technological advancements and the gradual maturation of the market, mRNA is set to play an increasingly significant role in versatile biotechnology fields.

Robust scale-up of bioreactor systems requires hydrodynamic similarity across scales to ensure consistent cell culture performance. This study presents a computational fluid dynamics (CFD)-guided digital framework for optimizing the scale-up of a novel orbital rocking bioreactor, CELBIC, from CELBIC5 (working volume 1–2 L) to CELBIC50 (working volume 10–20 L). Using lattice Boltzmann-based simulations, key hydrodynamic parameters, including velocity, shear stress, and energy dissipation rate, were evaluated across various working volumes, inclination angles, and agitation speeds. Compared with conventional scale-up criteria, such as average P/V, we propose a comprehensive and digitalized scale-up optimization framework: a root mean square error (RMSE)-based method comparing the full spatial and temporal distributions of various CFD variables to quantify similarities among scaled-up conditions. This approach allows identification of the best-matching scaled-up condition and further refinement using response surface analysis. The optimized condition (10 L, 7°, and 19 rpm) exhibited the lowest combined RMSE relative to the reference condition (CELBIC5 at 1 L, 6°, and 30 rpm), suggesting substantial hydrodynamic equivalence. Overall, this study demonstrates how integrating CFD- and RMSE-based analyses enables rational scale-up of orbital rocking bioreactors, offering a systematic strategy for digital process development in single-use systems.

Despite the challenge in generating adequate immunogenicity, peptide-based vaccines offer a significant benefit in terms of improving immunity when combined with conjugates. In previous studies, three computationally designed immunogenic peptides of the Zika prM protein were reported to induce an immune response. In the current work, reported peptides were conjugated with polymer, polyethylene glycol 400, with the intention of enhancing the immune response. Conjugation was confirmed by Fourier transform infrared spectroscopy. We reckoned the immune response of all three peptide-PEG conjugates (MPC1, MPC2, and MPC3) on 10 different healthy blood samples. Assessment of MPC-induced human peripheral blood mononuclear cell (PBMC) proliferation and interferon-gamma (IFN-γ) secretion was done with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and a sandwich enzyme-linked immunosorbent assay (ELISA). Conjugate MPC1 showed significantly enhanced cell proliferation in five and MPC2 and MPC3 in six samples, compared to the peptides. In case of IFN-γ release, conjugate MPC1 exhibited significant results in six, MPC2 in five, and MPC3 in seven samples elevated results, in contrast to peptides alone. Thus, conjugation of PEG to immunogenic peptides could be an effective way to increase the peptide immunogenicity, although further experimental validations are required before considering them as candidates for a vaccine against Zika virus infection.

Microfluidic brain-on-a-chip and angiogenesis-on-a-chip models have been employed by leading research groups around the world. These models have great potential, but they have not yet been used in the context of personalized medicine and diagnostics, nor have they been widely adopted for testing drug candidates. The reproduction of a physiologically relevant in vitro brain-on-a-chip model with essential similarity to in vivo structural and functional characteristics, including integrity and complexity of the brain tissue, limited and controlled permeability of barriers, and plastic changes induced by brain stimulation or injury, is a highly challenging task that requires a comprehensive approach and a tour de force in advanced biomedical engineering and living soft matter physics. This approach necessitates not only the combination of well-established chip production technologies and cell biology protocols but also the integration of knowledge from neuroscience, (bio)physics, hydrodynamics, material science, (bio)chemistry, electronics, and soft matter physics. This review covers current understandings of the establishment of microvascular network formation in the active brain in vivo, as well as analysis of novel approaches to reconstruct the basic mechanisms of cerebral angiogenesis and barrier genesis in a physiologically relevant brain-on-a-chip model. The novel concept, which employs the application of an external electric field to stimulate vasculogenesis/angiogenesis on a chip, demonstrates how a synergistic approach may help to solve this non-trivial problem and to establish the vascularized brain tissue sensitive to various stimuli and suitable for the high-precision analysis of its functional activity and plasticity.

One of the main obstacles to preparing oligoxanthan through the enzymatic degradation of xanthan gum (XG) is its dense structure. To achieve the large-scale production of oligoxanthan, it is necessary to perturbate the conformation of XG and increase enzyme accessibility. This research developed an efficient system for the enzymatic degradation of XG assisted by low concentrations of ionic liquids (ILs). Structural characterization results indicated that a low concentration of [EMIM]DEP (1%, v/v) not only loosens the structure of XG but also alters the secondary structure of enzyme, thereby dramatically increasing its hydrolytic activity and eventually enhancing the production of oligoxanthan. The results were further elucidated by molecular dynamics (MD) simulation. Under the hydrolysis conditions of 1.5 mg/mL Aspergillus niger cellulase (AnCel), 0.2 mg/mL XG, and 1% (v/v) [EMIM]DEP at 40°C and pH 5.0 for 20 min, XG can be hydrolyzed into a mixture with a glucose equivalent of 32.25%. The degree of polymerization (DP) of XG hydrolysates was 3–10. Moreover, the xanthan hydrolysates obtained using this system exhibit excellent antioxidant activity. This study proposed an effective method for the preparation of functional oligosaccharides, which is of significant importance for the valorization of XG.

Methylene-tetrahydrofolate reductase (MTHFR) is an important enzyme for acetogenic carbon fixation, but the redox mechanism driving this reaction is not clearly understood. Previous enzymology work and energetic accounting on species such as Clostridium autoethanogenum has led to confounding results when placed in the context of in vivo experiments. In this work, we create multiple C. autoethanogenum strains harboring alternative MTHFR enzyme complexes as well as genome-scale metabolic models to better understand how these organisms conserve energy on gas substrates. The inclusion of a Type-III MTHFR unexpectedly allows for higher growth than expected and suggests the possibility of an additional redox balancing cycle employed during autotrophic growth.