Engineering in Life Sciences最新文献

{"title":"Risk control of host cell proteins in one therapeutic antibody produced by concentrated fed-batch (CFB) mode","authors":"Yiling Lu,&nbsp;Jun Lin,&nbsp;Tianze Bian,&nbsp;Jin Chen,&nbsp;Dan Liu,&nbsp;Mingjun Ma,&nbsp;Zhen Gao,&nbsp;Jiemin Chen,&nbsp;Dianwen Ju,&nbsp;Xing Wang","doi":"10.1002/elsc.202200060","DOIUrl":"10.1002/elsc.202200060","url":null,"abstract":"<p>Multiple control strategies, including a downstream purification process with well-controlled parameters and a comprehensive release or characterization for intermediates or drug substances, were implemented to mitigate the potential risk of host cell proteins (HCPs) in one concentrated fed-batch (CFB) mode manufactured product. A host cell process specific enzyme-linked immunosorbent assay (ELISA) method was developed for the quantitation of HCPs. The method was fully validated and showed good performance including high antibody coverage. This was confirmed by 2D Gel-Western Blot analysis. Furthermore, a LC-MS/MS method with non-denaturing digestion and a long gradient chromatographic separation coupled with data dependent acquisition (DDA) on a Thermo/QE-HF-X mass spectrometer was developed as an orthogonal method to help identify the specific types of HCPs in this CFB product. Because of the high sensitivity, selectivity and adaptability of the new developed LC-MS/MS method, significantly more species of HCP contaminants were able to be identified. Even though high levels of HCPs were observed in the harvest bulk of this CFB product, the development of multiple processes and analytical control strategies may greatly mitigate potential risks and reduce HCPs contaminants to a very low level. No high-risk HCP was identified and the total amount of HCPs was very low in the CFB final product.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"23 3","pages":""},"PeriodicalIF":2.7,"publicationDate":"2023-02-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202200060","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10277423","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"An optimized live bacterial delivery vehicle safely and efficaciously delivers bacterially transcribed therapeutic nucleic acids","authors":"Darcy S. O. Mora,&nbsp;Madeline Cox,&nbsp;Forgivemore Magunda,&nbsp;Ashley B. Williams,&nbsp;Lyndsey Linke","doi":"10.1002/elsc.202200037","DOIUrl":"10.1002/elsc.202200037","url":null,"abstract":"<p>There is an unmet need for delivery platforms that realize the full potential of next-generation nucleic acid therapeutics. The in vivo usefulness of current delivery systems is limited by numerous weaknesses, including poor targeting specificity, inefficient access to target cell cytoplasm, immune activation, off-target effects, small therapeutic windows, limited genetic encoding and cargo capacity, and manufacturing challenges. Here we characterize the safety and efficacy of a delivery platform comprising engineered live, tissue-targeting, non-pathogenic bacteria (<i>Escherichia coli</i> SVC1) for intracellular cargo delivery. SVC1 bacteria are engineered to specifically bind to epithelial cells via a surface-expressed targeting ligand, to allow escape of their cargo from the phagosome, and to have minimal immunogenicity. We describe SVC1's ability to deliver short hairpin RNA (shRNA), localized SVC1 administration to various tissues, and its minimal immunogenicity. To validate the therapeutic potential of SVC1, we used it to deliver influenza-targeting antiviral shRNAs to respiratory tissues in vivo. These data are the first to establish the safety and efficacy of this bacteria-based delivery platform for use in multiple tissue types and as an antiviral in the mammalian respiratory tract. We expect that this optimized delivery platform will enable a variety of advanced therapeutic approaches.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"23 3","pages":""},"PeriodicalIF":2.7,"publicationDate":"2023-02-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202200037","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10837719","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Rapid in-situ aerobic biodegradation of high salt and oily food waste employing constructed synthetic microbiome","authors":"Song Xu,&nbsp;Lidan Tao,&nbsp;Jingjing Wang,&nbsp;Xiaoxia Zhang,&nbsp;Zhiyong Huang","doi":"10.1002/elsc.202200067","DOIUrl":"10.1002/elsc.202200067","url":null,"abstract":"<p>The high salt content of food waste (FW) severely limits microbial physiological activity and reduces its biodegradability. In this study, a salt-tolerant thermophilic bacterial agent that consists of four different substrate degradation functional strains was evaluated for efficient high salt and oily FW in solid-state aerobic biodegradation disposers. The phy-chemical properties, enzyme activities, microbial community structure, and function during the biodegradation process were evaluated under high salt (5%) stress. The results showed that the agent promoted the degradation rate, increased the matrix temperature, decreased the moisture content (MC), and enhanced enzyme activities without putrid smell. High-throughput sequencing indicated community structure succession between different groups and the positive contribution of the inoculated functional strains. During the FW biodegradation process, the <i>Bacillus</i> sp. inoculated was the dominant genus in the agent group. Furthermore, CCA further confirmed the positive effects of the four inoculated strains on high salt and oily FW aerobic biodegradation. Functional prediction and metabolite results both confirmed that the agent was more efficient in carbon, amino acid, and lipid metabolism, which demonstrated that the synthetic microbial consortium holds a potential advantage for efficiency and subsequent resource utilization for organic fertilizer.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"24 5","pages":""},"PeriodicalIF":2.7,"publicationDate":"2023-02-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202200067","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43104174","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cover Picture: Engineering in Life Sciences 2'23","authors":"","doi":"10.1002/elsc.202370021","DOIUrl":"https://doi.org/10.1002/elsc.202370021","url":null,"abstract":"","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"23 2","pages":""},"PeriodicalIF":2.7,"publicationDate":"2023-02-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202370021","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50120945","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Escherichia coli aceE variants coding pyruvate dehydrogenase improve the generation of pyruvate-derived acetoin","authors":"W. Chris Moxley,&nbsp;Rachel E. Brown,&nbsp;Mark A. Eiteman","doi":"10.1002/elsc.202200054","DOIUrl":"https://doi.org/10.1002/elsc.202200054","url":null,"abstract":"<p>Several chromosomally expressed AceE variants were constructed in <i>Escherichia coli ΔldhA ΔpoxB ΔppsA</i> and compared using glucose as the sole carbon source. These variants were examined in shake flask cultures for growth rate, pyruvate accumulation, and acetoin production via heterologous expression of the <i>budA</i> and <i>budB</i> genes from <i>Enterobacter cloacae ssp. dissolvens</i>. The best acetoin-producing strains were subsequently studied in controlled batch culture at the one-liter scale. PDH variant strains attained up to four-fold greater acetoin than the strain expressing the wild-type PDH. In a repeated batch process, the H106V PDH variant strain attained over 43 g/L of pyruvate-derived products, acetoin (38.5 g/L) and 2R,3R-butanediol (5.0 g/L), corresponding to an effective concentration of 59 g/L considering the dilution. The acetoin yield from glucose was 0.29 g/g with a volumetric productivity of 0.9 g/L·h (0.34 g/g and 1.0 g/L·h total products). The results demonstrate a new tool in pathway engineering, the modification of a key metabolic enzyme to improve the formation of a product via a kinetically slow, introduced pathway. Direct modification of the pathway enzyme offers an alternative to promoter engineering in cases where the promoter is involved in a complex regulatory network.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"23 3","pages":""},"PeriodicalIF":2.7,"publicationDate":"2023-01-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202200054","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50126325","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cell-free synthesis of silver nanoparticles in spent media of different Aspergillus species","authors":"Alexander Boldt,&nbsp;Jan Walter,&nbsp;Fabian Hofbauer,&nbsp;Karen Stetter,&nbsp;Ines Aubel,&nbsp;Martin Bertau,&nbsp;Christof M. Jäger,&nbsp;Thomas Walther","doi":"10.1002/elsc.202200052","DOIUrl":"https://doi.org/10.1002/elsc.202200052","url":null,"abstract":"<p>The recovery and valorization of metals and rare earth metals from wastewater are of great importance to prevent environmental pollution and recover valuable resources. Certain bacterial and fungal species are capable of removing metal ions from the environment by facilitating their reduction and precipitation. Even though the phenomenon is well documented, little is known about the mechanism. Therefore, we systematically investigated the influence of nitrogen sources, cultivation time, biomass, and protein concentration on silver reduction capacities of cell-free cultivation media (spent media) of <i>Aspergillus niger</i>, <i>A. terreus</i>, and <i>A. oryzae</i>. The spent medium of <i>A. niger</i> showed the highest silver reduction capacities with up to 15 μmol per milliliter spent medium when ammonium was used as the sole N-source. Silver ion reduction in the spent medium was not driven by enzymes and did not correlate with biomass concentration. Nearly full reduction capacity was reached after 2 days of incubation, long before the cessation of growth and onset of the stationary phase. The size of silver nanoparticles formed in the spent medium of <i>A. niger</i> was influenced by the nitrogen source, with silver nanoparticles formed in nitrate or ammonium-containing medium having an average diameter of 32 and 6 nm, respectively.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"23 3","pages":""},"PeriodicalIF":2.7,"publicationDate":"2023-01-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202200052","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50129832","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Using the inner membrane of Escherichia coli as a scaffold to anchor enzymes for metabolic flux enhancement","authors":"You Wang,&nbsp;Yushu Wang,&nbsp;Yuqi Wu,&nbsp;Yang Suo,&nbsp;Huaqing Guo,&nbsp;Yineng Yu,&nbsp;Ruonan Yin,&nbsp;Rui Xi,&nbsp;Jiajie Wu,&nbsp;Nan Hua,&nbsp;Yuehan Zhang,&nbsp;Shaobo Zhang,&nbsp;Zhenming Jin,&nbsp;Lin He,&nbsp;Gang Ma","doi":"10.1002/elsc.202200034","DOIUrl":"https://doi.org/10.1002/elsc.202200034","url":null,"abstract":"<p>Clustering enzymes in the same metabolic pathway is a natural strategy to enhance productivity. Synthetic protein, RNA and DNA scaffolds have been designed to artificially cluster multiple enzymes in the cell, which require complex construction processes and possess limited slots for target enzymes. We utilized the <i>Escherichia coli</i> inner cell membrane as a native scaffold to cluster four fatty acid synthases (FAS) and achieved to improve the efficiency of fatty acid synthesis in vivo. The construction strategy is as simple as fusing target enzymes to the N-terminus or C-terminus of the membrane anchor protein (Lgt), and the number of anchored enzymes is not restricted. This novel device not only presents a similar efficiency in clustering multiple enzymes to that of other artificial scaffolds but also promotes the product secretion, driving the entire metabolic flux forward and further increasing the gross yield compared with that in a cytoplasmic scaffold system.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"23 2","pages":""},"PeriodicalIF":2.7,"publicationDate":"2023-01-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202200034","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50127267","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Editorial for the special issue ‘cell-to-cell’ and ‘cell-to-bioreactor’ interactions","authors":"Ralf Takors","doi":"10.1002/elsc.202200062","DOIUrl":"10.1002/elsc.202200062","url":null,"abstract":"<p>Shifting the current fossil economy to a carbon neutral supply is an enormous challenge. Current geopolitical issues that severely endanger long-term established fossil supply chains function as catalysts fostering novel solutions. In this context, the goals for establishing a resilient economy meet climate protection demands for reducing the human footprint in atmosphere. Biotechnological processes for producing fine chemicals and commodities by utilizing renewable resources may play an important role to make the transformation to a circular economy happen.</p><p>Bioprocesses already successfully documented their outstanding quality not only to compete with established fossil routes but also to complement and even to replace them in industrial practice. Long-term established examples comprise the microbial production of amino acids, organic acids, technical enzymes, food additives, active pharma ingredients … and many more.</p><p>Interestingly enough, mono-cultures are used, predominately. This reflects the steady improvement of molecular tools for efficiently manipulating microbes to produce the molecule of interests. However, the question may arise whether mono-cultures should be the first choice for developing novel bioprocesses. Often enough product formation demands for precursors, reduction equivalents, energy demands, etc. that contradict cellular needs for growth and maintenance. Furthermore, the genetic engineering of hosts may well reach technical limits if targeted product formation opposes the lifestyle of the cells. Additionally, many examples for the production of antibiotics are known outlining that antibiotic production only starts in the presence of another interacting strain.</p><p>Consequently, the implementation of co-cultures with well-equilibrated interactions is a promising approach for establishing a new generation of bioproduction processes. Accordingly, this topic (cell-to-cell interactions) is highlighted in the current special issue and is also a central theme in the priority program ‘InterZell SPP2170’ of the German Science Foundation (DFG) that co-fuels the special issue.</p><p>Once developed in the labs, novel bioprocesses should find their way into large-scale bioreactors to translate innovation into practice. Often enough the so-called scale-up reveals the deterioration of key performance criteria such as titer, rates, and yield (TRY). This basically reflects cellular responses on mixing heterogeneities that inevitably occur in industrial scale. For mitigating related performance losses profound research is necessary. Consequently, the special issue also covers related studies (cell-to-bioreactor interactions) analyzing microbial responses in detail and developing novel scale-down devices.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"23 1","pages":""},"PeriodicalIF":2.7,"publicationDate":"2023-01-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202200062","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10497824","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cover Picture: Engineering in Life Sciences 1'23","authors":"","doi":"10.1002/elsc.202370011","DOIUrl":"https://doi.org/10.1002/elsc.202370011","url":null,"abstract":"","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"23 1","pages":""},"PeriodicalIF":2.7,"publicationDate":"2023-01-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202370011","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50120749","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Mobile SARS‑CoV‑2 screening facilities for rapid deployment and university-based diagnostic laboratory","authors":"Nils Stanislawski,&nbsp;Ferdinand Lange,&nbsp;Christian Fahnemann,&nbsp;Christoph Riggers,&nbsp;Marc-Nils Wahalla,&nbsp;Marc Porr,&nbsp;Fabian Cholewa,&nbsp;Rebecca Jonczyk,&nbsp;Stefanie Thoms,&nbsp;Martin Witt,&nbsp;Frank Stahl,&nbsp;Sascha Beutel,&nbsp;Andreas Winkel,&nbsp;Philipp-Cornelius Pott,&nbsp;Meike Stiesch,&nbsp;Mira Paulsen,&nbsp;Anette Melk,&nbsp;Henning Lucas,&nbsp;Stefanie Heiden,&nbsp;Holger Blume,&nbsp;Cornelia Blume","doi":"10.1002/elsc.202200026","DOIUrl":"10.1002/elsc.202200026","url":null,"abstract":"<p>The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has created a public crisis. Many medical and public institutions and businesses went into isolation in response to the pandemic. Because SARS-CoV-2 can spread irrespective of a patient's course of disease, these institutions’ continued operation or reopening based on the assessment and control of virus spread can be supported by targeted population screening. For this purpose, virus testing in the form of polymerase chain reaction (PCR) analysis and antibody detection in blood can be central. Mobile SARS-CoV-2 screening facilities with a built-in biosafety level (BSL)-2 laboratory were set up to allow the testing offer to be brought close to the subject group's workplace. University staff members, their expertise, and already available equipment were used to implement and operate the screening facilities and a certified diagnostic laboratory. This operation also included specimen collection, transport, PCR and antibody analysis, and informing subjects as well as public health departments. Screening facilities were established at different locations such as educational institutions, nursing homes, and companies providing critical supply chains for health care. Less than 4 weeks after the first imposed lockdown in Germany, a first mobile testing station was established featuring a build-in laboratory with two similar stations commencing operation until June 2020. During the 15-month project period, approximately 33,000 PCR tests and close to 7000 antibody detection tests were collected and analyzed. The presented approach describes the required procedures that enabled the screening facilities and laboratories to collect and process several hundred specimens each day under difficult conditions. This report can assist others in establishing similar setups for pandemic scenarios.</p>","PeriodicalId":11678,"journal":{"name":"Engineering in Life Sciences","volume":"23 2","pages":""},"PeriodicalIF":2.7,"publicationDate":"2023-01-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/elsc.202200026","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10666277","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}