Biotechnology Reports最新文献

Human induced pluripotent stem cells (iPSC) have demonstrated massive potentials for use in regenerative and personalized medicine due to their ability to expand in culture and differentiate into specialized cells with therapeutic benefits. However, in order to industrialize iPSC-derived therapies, it is necessary to address the existing challenges surrounding the analytics implemented in the manufacturing process to evaluate and monitor cell expansion, differentiation, and quality of the final products. Here, we review some of the key analytical methods used as part of identity, potency, or safety for in-process or final product release testing and highlighted the challenges and potential solutions for consideration in the Chemistry, Manufacturing and Controls (CMC) strategy for iPSC-based therapies.

Some of the challenges associated with characterization and testing of iPSC-based products are related to the choice of analytical technology (to ensure fit-for-purpose), assay reliability and robustness. Automation of analytical methods may be required to reduce hands on time, and improve reliability of the methods through reducing assay variability. Indeed, we have shown that automation of analytical methods is feasible (evaluated using an ELISA based assay) and would result in more precise measurements (demonstrated by lower co-efficient of Variation and standard deviation), less hands-on time, and swift compared to a manually run assay. Therefore, in order to support commercialization of iPSC-based therapies we suggest a well-designed testing strategy to be established in the development phase while incorporating robust, reproducible, reliable, and potentially automated analytics in the manufacturing process.

Marine macroalgae are being recognized as reservoirs of biologically active compounds, as their surfaces are susceptible to the colonization of microorganisms which can produce enzymes with a wide range of molecular architectures. Among these bacteria, Achromobacter is responsible for the biosynthesis of laccases. In this research, we performed a bioinformatic pipeline to annotate the sequenced complete genome of the epiphytic bacterium Achromobacter denitrificans strain EPI24, from the macroalgal surface of the Ulva lactuca species; this strain showed laccase activity which has been previously assessed on plate assays. The genome of A. denitrificans strain EPI24 has a size of ∼6.95 Mb, a GC content of 67.33%, and 6,603 protein-coding genes. The functional annotation of the A. denitrificans strain EPI24 genome confirmed the presence of genes encoding for laccases, which could have functional properties of interest in processes such as the biodegradation of phenolic compounds under versatile and efficient conditions.

This work aimed to carry out chemical cooking of corn stalks, both in a nitrate-alkaline manner and in a soda pulp method. The composition of corn is characterized by cellulose, lignin, ash, and substances extractable into polar and organic solvents. Handsheets were made from the pulp, for which the degree of polymerization, sedimentation rate, and strength properties was determined.

SARS-CoV-2 receptor binding domain (RBD) recognizes the angiotensin converting enzyme 2 (ACE2) receptor in host cells that enables infection. Due to its antigenic specificity, RBD production is important for development of serological assays. Here we have established a system for stable RBD production in HEK 293T mammalian cells that simultaneously express the recombinant fluorescent protein dTomato, which enables kinetic monitoring of RBD expression by fluorescence microscopy. In addition, we have validated the use of this recombinant RBD in an ELISA assay for the detection of anti-RBD antibodies in serum samples of COVID-19 convalescent patients. Recombinant RBD generated using this approach can be useful for generation of antibody-based therapeutics against SARS-CoV-2, as well serological assays aimed to test antibody responses to this relevant virus.

Seed coating is considered one of the best methods to promote sustainable agriculture where the physical and physiological properties of seeds can be improved to facilitate planting, increase growth indices and alleviate abiotic and biotic stresses. Several methods of seed coating are used to attain good application uniformity and adherence in the seed coating process. Seed coating has been tested in seeds of various plant species with different dimensions, forms, textures, and germination types. Plant beneficial microorganisms (PBM), such as rhizobia, bacteria, and fungi inoculated via seed inoculation can increase seed germination, plant performance and tolerance across biotic (e.g., pathogens and pests) and abiotic stress (e.g., salt, drought, and heavy metals) while reducing the use of agrochemical inputs. In this review, the microbial seed coating process and their ability to increase seed performance and protect plants from biotic and abiotic stresses are well discussed and highlighted in sustainable agricultural systems.

The outbreak of the novel severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that causes the Coronavirus Disease 2019 (COVID-19) has spread through the globe at an alarming speed. The disease has become a global pandemic affecting millions of people and created public health crises worldwide. Among many efforts to urgently develop a vaccine against this disease, we developed an industrial-scale closed, single use manufacturing process for V590, a vaccine candidate for SARS-CoV-2. V590 is a recombinant vesicular stomatitis virus (rVSV) genetically engineered to express SARS-CoV-2 glycoprotein. In this work, we describe the development and optimization of serum-free microcarrier production of V590 in Vero cells in a closed system. To achieve the maximum virus productivity, we optimized pH and temperature during virus production in 3 liters (L) bioreactors. Virus productivity was improved (by ∼1 log) by using pH 7.0 and temperature at 34.0 °C. The optimal production condition was successfully scaled up to a 2000 L Single Use Bioreactor (SUB), producing a maximum virus titer of ∼1.0e+7 plaque forming units (PFU)/mL. Further process intensification and simplification, including growing Vero cells at 2 gs per liter (g/L) of Cytodex-1 Gamma microcarriers and eliminating the media exchange (MX) step prior to infection helped to increase virus productivity by ∼2-fold.

Selenium nanoparticles (SeNPs) have antimicrobial and antifungal activity. SeNPs using Se resistant bacteria is a low cost and eco-friendly technology. Fungal contamination of wood during drying is one of the main causes of economic losses in the wood industry. The bacterium Delftia sp. 5 resistance to Se and its ability to produce SeNPs able to inhibit the growth of the wood brown-rotting fungus Oligoporus pelliculosus was analyzed. The strain showed an optimal SeNPs production when selenite concentration was 160 mg L⁻¹. The SeNPs were spherical with an average size 192.33 ± 8.6 nm and a zeta potential of -41.4 ± 1.3 nm. The SeNPs produced by Delftia sp. 5 (33.6 ± 0.1 mg L⁻¹ Se) inhibited the growth of O. pelliculosus in agar plates and in Nothofagus pumilio (Lenga) wood samples. Delftia sp. 5 SeNPs could be used for embedding lenga wood prior to drying for preventing the growth of the deteriorating fungi O. pelliculosus.

SARS-CoV-2 causes devastating impact on the human population and has become a major public health concern. The frequent emergence of SARS-CoV-2 variants of concern urges the development of safe and efficacious vaccine against SARS-CoV-2 variants. We developed a candidate vaccine Baiya SARS-CoV-2 Vax 1, based on SARS-CoV-2 receptor-binding domain (RBD) by fusing with the Fc region of human IgG. The RBD-Fc fusion was produced in Nicotiana benthamiana. Previously, we reported that this plant-produced vaccine is effective in inducing immune response in both mice and non-human primates. Here, the efficacy of our vaccine candidate was tested in Syrian hamster challenge model. Hamsters immunized with two intramuscular doses of Baiya SARS-CoV-2 Vax 1 induced neutralizing antibodies against SARS-CoV-2 and protected from SARS-CoV-2 challenge with reduced viral load in the lungs. These preliminary results demonstrate the ability of plant-produced subunit vaccine Baiya SARS-CoV-2 Vax 1 to provide protection against SARS-CoV-2 infection in hamsters.

Sunburn is one of the most common skin lesions caused by excessive UV exposure, and its incidence is highly correlated with the risks of skin cancer. A variety of drugs including corticosteroids and NSAIDs have been developed to treat acute sunburn, however, they have raised severe concerns such as poor healing efficacy and long recovery time. We have for the first time extracted non-denatured type I collagen from yak hide, which displays a canonical triple helical structure with melting temperature of 42.7 °C. The highly pure yak collagen type I (YCI) self-assembles to form well-ordered nanofibers with periodic d-bands. YCI is highly biocompatible, and it significantly promotes the proliferation and adhesion of HFF-1 cells. The sunburn healing effects of YCI has been investigated using acute skin injury mouse model. Histological analysis shows that 4 days’ treatment of YCI has resulted in the recovery of sunburned mice skin to a healthy state, indicated by pronounced acceleration of epithelization and collagen deposition. The collagen volume fraction as well as the hydroxyproline (Hyp) content of YCI-treated sunburned skin have been found to be greatly increased, confirming the enhanced regeneration of collagen. YCI creams and dressings have also shown superior healing capacity of sunburn by remarkably shortening the recovery time. Notably, the denatured collagen-targeted staining results indicated a large quantity of denatured collagen in sunburned mice, which became substantially reduced after the YCI treatment. FITC-labeled YCI has been further found to penetrate into the dermis of sunburned mice. The highly biocompatible and bioactive non-denatured YCI provides an improved treatment of sunburn, indicating very promising applications of YCI in cosmetics and dermatology.

The potential for the application of metal-containing nanomaterials at the nanoscale promotes the opportunity to search for new methods for their elaboration, with special attention to those sustainable methods. In response to these challenges, we have investigated a new method for green synthesis of cuprous oxide nanoparticles (Cu₂O NPs) using Myrciaria dubia juice as an organic reductant and, comparing it with chemical synthesis, evaluating in both cases the influence of the volume of the organic (juice) and chemical (ascorbic acid) reductants, for which a large number of techniques such as spectrophotometry, EDX spectrometry, TEM, SEM, DLS, FTIR spectroscopy have been used. Likewise, the nanomaterial with better morphological characteristics, stability, and size homogeneity has been applied in the functionalization of textiles by means of in situ and post-synthesis impregnation methods. The success of the synthesis process has been demonstrated by the antimicrobial activity (bacteria and fungi) of textiles impregnated with Cu₂O NPs.