Engineering in Life Sciences最新文献

Microplastics (MPs) are major concern due to their potential harm to ecosystems and most research has focused on their presence and fate, with limited attention to their biodegradation in aquatic ecosystems. Nevertheless, MPs act as hotspots for the colonization by a diverse range of microorganisms that can adhere to plastic surfaces, resulting in the subsequent formation of biofilms—a potential threat especially in terms of pathogenicity. This study employed 16S rRNA and 18S rRNA sequencing metagenomic analyses to investigate microbial communities within biofilms on plastic materials exposed to long-term marine and freshwater environments. Three Arcobacter species (Arcobacter nitrofigilis, Arcobacter acticola, and Arcobacter suis) emerged as dominant species in M_MP sample, while Flavobacterium tructae was the predominant species within the F_MP sample. The 18S rRNA sequencing revealed the presence of the fungal phylum Ascomycota and the microalgal species Pseudocharaciopsis ovalis in F_MP. Although, the primary species detected on M_MP and F_MP samples include bacteria previously implicated as pathogen, the predominant species identified in this study were unconnected to MP-associated biofilms or MP degradation. Their presence constitutes a novel discovery, opening promising avenues for the exploration of their potential involvement in the biodegradation of MPs within aquatic environments.

Polyhydroxyalkanoates (PHAs) have gained interest recently due to their biodegradability and versatility. In particular, the chemical compositions of medium-chain-length (mcl)-PHAs are highly diverse, comprising different monomers containing 6–14 carbon atoms. This review summarizes different feedstocks and fermentation strategies to enhance mcl-PHA production and briefly discusses the downstream processing. This review also provides comprehensive details on analytical tools for determining the composition and properties of mcl-PHA. Moreover, this study provides novel information by statistically analyzing the data collected from several reports on mcl-PHA to determine the optimal fermentation parameters (specific growth rate, PHA productivity, and PHA yield from various structurally related and unrelated substrates), mcl-PHA composition, molecular weight (MW), and thermal and mechanical properties, in addition to other relevant statistical values. The analysis revealed that the median PHA productivity observed in the fed-batch feeding strategy was 0.4 g L⁻¹ h⁻¹, which is eight times higher than that obtained from batch feeding (0.05 g L⁻¹ h⁻¹). Furthermore, 3-hydroxyoctanoate and -decanoate were the primary monomers incorporated into mcl-PHA. The investigation also determined the median glass transition temperature (−43°C) and melting temperature (47°C), which indicated that mcl-PHA is a flexible amorphous polymer at room temperature with a median MW of 104 kDa. However, information on the monomer composition or heterogeneity and the associated physical and mechanical data of mcl-PHAs is inadequate. Based on their mechanical values, the mcl-PHAs can be classified as semi-crystalline polymers (median crystallinity 23%) with rubber-like properties and a median elongation at break of 385%. However, due to the limited mechanical data available for mcl-PHAs with known monomer composition, identifying suitable processing tools and applications to develop mcl-PHAs further is challenging.

Innovations in viral vaccine manufacturing are crucial for pandemic preparedness and to meet ever-rising global demands. For influenza, however, production still mainly relies on technologies established decades ago. Although modern production shifts from egg-based towards cell culture technologies, the full potential has not yet been fully exploited. Here, we evaluate whether implementation of state-of-the-art technologies for cell culture-based recombinant protein production are capable to challenge outdated approaches in viral vaccine process development. For this, a fully automated single-cell cloning strategy was established to generate monoclonal suspension Madin-Darby canine kidney (MDCK) cells. Among selected cell clones, we could observe distinct metabolic and growth characteristics, with C59 reaching a maximum viable cell concentration of 17.3 × 10⁶ cells/mL and low doubling times in batch mode. Screening for virus production using a panel of human vaccine-relevant influenza A and B viruses in an ambr15 system revealed high titers with yields competing or even outperforming available MDCK cell lines. With C113, we achieved cell-specific virus yields of up to 25,000 virions/cell, making this cell clone highly attractive for vaccine production. Finally, we confirmed process performance at a 50-fold higher working volume. In summary, we present a scalable and powerful approach for accelerated development of high-yield influenza virus production in chemically defined medium starting from a single cell.

Digitalization with integrated devices, digital and physical assistants, automation, and simulation is setting a new direction for laboratory work. Even with complex research workflows, high staff turnover, and a limited budget some laboratories have already shown that digitalization is indeed possible. However, academic bioprocess laboratories often struggle to follow the trend of digitalization. Due to their diverse research circumstances, high variety of team composition, goals, and limitations the concepts are substantially different. Here, we will provide an overview on different aspects of digitalization and describe how academic laboratories successfully digitalized their working environment. The key aspect is the collaboration and communication between IT-experts and scientific staff. The developed digital infrastructure is only useful if it supports the laboratory worker and does not complicate their work. Thereby, laboratory researchers have to collaborate closely with IT-experts in order for a well-developed and maintainable digitalization concept that fits their individual needs and level of complexity. This review may serve as a starting point or a collection of ideas for the transformation toward a digitalized laboratory.

Process Analytical Technologies (PATs) are taking a key role in the run for automatization in the biopharmaceutical industry. Spectroscopic methods such as Raman spectroscopy or mid-infrared (MIR) spectroscopy are getting more recognition in the recent years for inline monitoring of bioprocesses due to their ability to measure various molecules simultaneously. However, their dependency on laborious model calibration making them a challenge to implement. In this study, a novel one-point calibration that requires a single reference point prior to the inline monitoring of glucose and lactate in bioprocesses with MIR spectroscopy is assessed with 22 mammalian cell perfusion (PER) processes in two different scales and four different products. Concentrations are predicted over all PERs runs with a root mean square error (RMSE) of 0.29 g/L for glucose and 0.24 g/L for lactate, respectively. For comparison conventional partial least square regression (PLSR) models were used and trained with spectroscopic data from six bioreactor runs in two different scales and three products. The general accuracy of those models (RMSE of 0.41 g/L for glucose and 0.16 g/L for lactate) are in the range of the accuracy of the one-point calibration. This shows the potential of the one-point calibration as an approach making spectroscopy more accessible for bioprocess development.