Engineering in Life Sciences最新文献

Autologous chimeric antigen receptor (CAR)-T therapies have given hope to many cancer patients whose other lines of treatment have failed. Unfortunately, limited manufacturing capability has resulted in many patients dying while on a waitlist. Similarly, since clinical trial treatments are personalized, it is difficult to treat many patients simultaneously, resulting in longer clinical trials. Therapeutic production often takes over 4 weeks, so a product failure means that a patient may need to wait another month for treatment, putting them at severe risk for disease progression. The labor-intensive manufacturing process has led to therapeutic costs of roughly $500,000 per treatment, which can be reduced by better automation and shorter manufacturing times. The goals of this article are to review CAR-T therapeutics development, manufacturing, and treatment, and to encourage the development of data analytics-based multi-scale decision support tools for all humans “in the loop.” A systems approach is needed since prior treatments and current state of health (including the immune system and microbiota), initial cell quality, manufacturing failure, bridging and lymphodepletion therapy before infusion, and supply chain management, all impact treatment success. Continuous updates as more patient data are made available can lead to better treatment recommendations and outcomes.

Biosensors are an integral part of modern medicine, are used in basic research, and are increasingly used by consumers as point-of-care and wearable devices. Meanwhile, the underlying technological approaches are rapidly expanding, including spectroscopic sensing, artificial bioreceptors, synthetic biological approaches, whole-cell biosensors, and artificial intelligence. With these diversifications in applications as well as technology, the scope and meaning of the term biosensor is blurring. This paper attempts to give an overview of the sensing approaches, with their physical, chemical, biochemical, and biological principles, and an overview of the fields of application, including nonliving systems and living systems. This leads to a comprehensive overview and a reappreciation of the term biosensor, including not only devices with a molecular biorecognition element and physico-chemical readout but also the sensing of living biological systems using physical and chemical methods, and the use of living biological systems for sensing purposes.

Tissue engineering, and in particular the development of organ-on-a-chip (OOC) models, holds significant promise for advancing personalized medicine and reducing the use of animal models. The integration of microfluidics and advanced biomaterials in OOC systems provides controlled microenvironments and fosters the creation of physiologically relevant tissue models. A critical aspect of OOC models is the fabrication of perfusable chips to create vascular networks that are essential for sustaining long-term 3D cultures. Here we show a two-step fabrication approach that combines one- and two-photon polymerization (2PP) to create a microfluidic chip capable of supporting endothelial cell (EC) angiogenesis. The chip features a 2PP-printed sealing contour to ensure leak-free bonding of chip parts, and an array of channel-separating-pillars that enable EC migration from the parent vessel into an extracellular matrix. Our results demonstrate that the developed angiogenesis-on-a-chip model successfully induces EC sprouting in response to angiogenic factors. This work significantly contributes to the field by providing a versatile platform for vascular studies, highlighting the potential for its application in drug screening. The flexibility and precision of our fabrication method also allows for customizing OOC devices for various biological applications, thereby enhancing the relevance of these systems in investigation of complex tissue interactions.

The COVID-19 pandemic has highlighted the need for rapid, simple, and cost-effective point-of-care testing (POCT) methods for pathogen detection. Hydrogel-based biosensing has emerged as an increasingly popular approach, offering advantages such as reagent storage and multiplexing capabilities. In this study, we have functionalized different polymer substrates to ensure an adequate adherence of methacryloyl-modified gelatin-based hydrogel spots that function as biosensors. Different hydrogel formulations were tested for their suitability in a point-of-care testchip. Our findings demonstrate the good adherence properties of amino-functionalized and subsequent methacrylated polymer materials, specifically polyethylene terephthalate (PET) and oriented polypropylene (OPP), when used as substrates for hydrogel biosensors. Moreover, we successfully identified an optimal formulation for the hydrogel ink, consisting of amino-functionalized methacryloyl-modified gelatin with a biopolymer content of 3.5% (w/w) and a photoinitiator content of 0.0875%. This formulation not only enables printing with a piezoelectric 2D printer but also exhibits excellent hydrogel stability and adherence to the functionalized substrates. These results contribute to the development of reliable and efficient POCT methods for pathogen detection, addressing the limitations of current diagnostic capabilities. The study emphasizes inkjet-based functionalization, with comprehensive characterization of printability including viscosity, surface tension, and density, and provides expanded methodological details to ensure reproducibility. Practical application: This study demonstrates the successful functionalization of PET and OPP polymers as substrates for point-of-care test chips, paving the way for advanced diagnostic solutions. By amino-functionalization and methacryloylation of the surfaces, covalent bonding with biosensors was achieved, ensuring stability and adherence. A methacryloyl-modified gelatin-based hydrogel ink, optimized for piezoelectric printing, was identified for biosensor fabrication. The selected ink minimizes fluorophore quenching, preserving biosensor sensitivity. Among the tested materials, OPP showed superior adherence due to its non-polar characteristics. These findings enable the creation of multiplexed test chips capable of detecting multiple pathogens simultaneously, addressing a crucial gap in rapid and reliable diagnostics. Although the hydrogel-based biosensors have not yet been tested with encapsulated LAMP, this integration marks the next step toward fully functional point-of-care testing. Ultimately, this research advances the development of robust diagnostic platforms, with applications in healthcare settings for timely pathogen identification and disease management.

Microbial synthesis using renewable C₁-carbon sources like electrochemically produced formate (e-formate) represents a promising approach for climate-neutral chemical production. This study investigates formate utilization for ectoine biosynthesis by the halophilic methylotroph Methyloligella halotolerans. Preliminary growth assays confirmed formate utilization using 15–20 mM formate as the sole energy source substrate, when supplemented with yeast extract or vitamin solutions in a mineral salt medium. In a systematic study for ectoine production, formate utilization reached 0.305 ± 0.020 mmol d⁻¹ at 20 mM. With different C₁-substrates at 20 mM (3 mmol), ectoine production reached 10.3 ± 3.2 µmol (from methanol), 6.5 ± 0.8 µmol (from equimolar methanol/formate mix), 4.4 ± 0.1 µmol (from formate), and 1.2 ± 0.1 µmol (from e-formate). Medium buffering, pH stability and toxicity limited performance when formate and e-formate were supplied. Although ectoine yields were suboptimal, the feasibility of e-formate-based ectoine biosynthesis under high-salinity conditions with 9% NaCl, as shown in this study, discloses the great potential for integrating highly efficient electrochemical CO₂ reduction in saline media with microbial synthesis of organic chemicals.

Combining the volumetric fabrication of hydrogel constructs using extrusion bioprinting with highly precise drop-on-demand (DoD) bioprinting offers exciting opportunities in biofabrication. This technical report presents a technique in which a solenoid micro-pipette is operated as an additional tool in an extrusion (bio)printing system to deposit small volumes of bioinks into extrusion-printed hydrogel constructs. Using three exemplary approaches, we show that this enables the patterned placement of cells or growth factors within 3D constructs and thus influences developmental processes. Human cells within low-viscosity bioinks, deposited into extrusion-printed hydrogel constructs by filling inter-strand cavities or by injection into the hydrogel strands, maintained their viability and functionality up to 28 days. As demonstrated for salivary gland cells, the properties of the hydrogel matrix can influence the fate of the injected cells: In a stiff alginate (Alg)-based hydrogel, they formed aggregates, which is beneficial for organoid formation, and in softer hydrogels, they migrated to neighboring cell clusters. Locally injected signaling factors such as vascular endothelial growth factor (VEGF) attracted endothelial cells and fibroblasts, which migrated into previously cell-free hydrogel areas. The combination of extrusion and DoD bioprinting opens new approaches to integrate different cell types and functionalizations in one construct, facilitating the creation of more complex and dynamic models.

Fungal cell disruption plays a critical role in unlocking a wide range of high-value intracellular products such as lipids, proteins, pigments, and bioactive compounds. However, lysing fungal cells is far more challenging than breaking bacterial or algal cells due to their robust and highly structured cell walls. These biological barriers demand a tailored and strategic approach depending on the fungal species, cell morphology, and downstream processing requirements. This review explores the various mechanical and non-mechanical methods used to disrupt fungal cells, beyond outlining the core principles behind each method, the engineering and process factors that influence their performance are emphasized. A comparative analysis is provided, focusing on key parameters like disruption efficiency, scalability, cost-effectiveness, and environmental impact. The review also sheds light on emerging hybrid and integrated approaches, the role of pre-treatment or co-treatment strategies, and the potential for greener and more sustainable alternatives aligned with circular bioeconomy goals. Ultimately, this review aims to serve as a guide for researchers, bioprocess engineers, and industry professionals seeking to optimize fungal bioproduct extraction in a way that is not only technically sound but also economically viable and environmentally responsible, paving the way for more efficient, scalable, and sustainable fungal-based biomanufacturing.

Today, most recombinant protein drugs are produced by mammalian cells in a stirred-type bioreactor (BR). Although cell culture scale-up strategies have been extensively investigated, scale-up and switching BRs while maintaining comparable culture performance remains a challenging step. This is because the empirical correlations used to determine operating parameters are applicable only for limited situations using similar BRs across scales. In addition, a few small scale-down models (SSDMs) are able to evaluate cellular sensitivity to the shear environment of manufacturing-scale BRs. In this study, we focused on the hydrodynamic stress associated with agitation and developed an SSDM that generates high shear stress without undesirable secondary effects such as vortex formation and severe gas hold-up. In-house BRs with various scales and configurations were used for fed-batch culture of CHO-K1 cells, and their shear environment was characterized by computational fluid dynamics (CFD). Using the dry-wet approach, we found that average shear stress was well correlated with titer decrease as an indicator of culture performance. We also confirmed that the response to shear stress differs among cell lines, and that evaluation of the shear sensitivity of cells is accordingly a risk mitigation step that is required to ensure successful scale-up.

Sequencing depth is a crucial parameter for variant calling accuracy and sensitivity. The trade-off between sequencing breadth and depth is a well-known limitation in capture-based targeted next-generation sequencing (NGS). Herein, we propose a differential depth sequencing method, SPRE-Seq, to acquire different sequencing depths for different targeted regions in an NGS panel. The SPRE-Seq performance was evaluated using a panel of reference standards and clinical samples based on our custom-designed homologous recombination deficiency (HRD) assay. By applying SPRE-Seq, the effective sequencing depths of the homologous recombination repair (HRR) and HRD regions of all seven HRD reference standards met the required thresholds with only half the sequencing data volume (reduced from 12 to 6 GB). The results for the HRR genes and HRD showed 100% consistency with the expected results. In clinical samples, the effective sequencing depth of the HRR regions was significantly higher, with a sequencing data volume of 6 GB using the SPRE-Seq approach compared with 6 GB using a regular capture approach. However, there was no significant difference between a data volume of 6 GB using SPRE-Seq and 12 GB using a regular capture method. The SPRE-Seq approach was feasible and reliable for determining the HRD status and HRR somatic variants in reference standards and clinical samples at a low sequencing volume. SPRE-Seq is a reliable, feasible, and cost-effective method that can acquire an adequate sequencing depth of an NGS panel at a low sequencing data volume.