Biotechnology and Bioengineering最新文献

CILF: CRISPR/Cas9 based integration of large DNA fragments in Saccharomyces cerevisiae. CILF：基于 CRISPR/Cas9 技术在酿酒酵母中整合大 DNA 片段。

IF 3.5 2区生物学

Biotechnology and Bioengineering Pub Date : 2024-12-01 Epub Date: 2024-08-18 DOI: 10.1002/bit.28830

Shijie Xu, Jie Meng, Qi Zhang, Baisong Tong, Zihe Liu, Jinyu Fu, Shuobo Shi

引用次数: 0

Sequential delivery of photosensitizers and checkpoint inhibitors by engineered bacteria for enhanced cancer photodynamic immunotherapy. 通过工程细菌依次递送光敏剂和检查点抑制剂，增强癌症光动力免疫疗法。

IF 3.5 2区生物学

Biotechnology and Bioengineering Pub Date : 2024-12-01 Epub Date: 2024-08-13 DOI: 10.1002/bit.28829

Xinyu Liu, Yali Fan, Xinyu Zhang, Lianyue Li, Chao Yang, Xiaoyan Ma, Guijie Bai, Dawei Sun, Yaxin Wang, Junyi Wang, Yong Li, Yanyan Shi, Jing Liu, Yingying Zhang, Hanjie Wang

{"title":"Sequential delivery of photosensitizers and checkpoint inhibitors by engineered bacteria for enhanced cancer photodynamic immunotherapy.","authors":"Xinyu Liu, Yali Fan, Xinyu Zhang, Lianyue Li, Chao Yang, Xiaoyan Ma, Guijie Bai, Dawei Sun, Yaxin Wang, Junyi Wang, Yong Li, Yanyan Shi, Jing Liu, Yingying Zhang, Hanjie Wang","doi":"10.1002/bit.28829","DOIUrl":"10.1002/bit.28829","url":null,"abstract":"Engineered bacteria-based cancer therapy has increasingly been considered to be a promising therapeutic strategy due to the development of synthetic biology. Wherein, engineering bacteria-mediated photodynamic therapy (PDT)-immunotherapy shows greater advantages and potential in treatment efficiency than monotherapy. However, the unsustainable regeneration of photosensitizers (PSs) and weak immune responses limit the therapeutic efficiency. Herein, we developed an engineered bacteria-based delivery system for sequential delivery of PSs and checkpoint inhibitors in cancer PDT-immunotherapy. The biosynthetic pathway of 5-aminolevulinic acid (5-ALA) was introduced into Escherichia coli, yielding a supernatant concentration of 172.19 mg/L after 10 h of growth. And another strain was endowed with the light-controllable releasement of anti-programmed cell death-ligand 1 nanobodies (anti-PD-L1). This system exhibited a collaborative effect, where PDT initiated tumor cell death and the released tumor cell fragments stimulated immunity, followed by the elimination of residual tumor cells. The tumor inhibition rate reached 74.97%, and the portion of activated T cells and inflammatory cytokines were reinforced. The results demonstrated that the engineered bacteria-based collaborative system could sequentially deliver therapeutic substance and checkpoint inhibitors, and achieve good therapeutic therapy. This paper will provide a new perspective for the cancer PDT-immunotherapy.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":null,"pages":null},"PeriodicalIF":3.5,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141975065","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Systematic mutational analysis reveals an essential role of N275 in IgE stability. 系统突变分析揭示了 N275 在 IgE 稳定性中的重要作用。

IF 3.5 2区生物学

Biotechnology and Bioengineering Pub Date : 2024-12-01 Epub Date: 2024-08-20 DOI: 10.1002/bit.28826

Shikha Kumari, Sanjay Ghosh, Saurabh Joshi, Ralf Guenther, Vanessa Siegmund, Achim Doerner

{"title":"Systematic mutational analysis reveals an essential role of N275 in IgE stability.","authors":"Shikha Kumari, Sanjay Ghosh, Saurabh Joshi, Ralf Guenther, Vanessa Siegmund, Achim Doerner","doi":"10.1002/bit.28826","DOIUrl":"10.1002/bit.28826","url":null,"abstract":"Therapeutic antibodies have predominantly been IgG-based. However, the ongoing clinical trial of MOv18 IgE has highlighted the potential of using IgE antibodies in cancer therapy. While extensive studies targeting IgG glycosylation resulted in a rational basis for the development of enhanced biotherapeutics, IgE glycosylation remains an area with limited analyses. Previous studies on the role of IgE glycosylation present conflicting data with one study emphasizing the importance of N275 and T277 residues for FcεRI binding whereas another asserts the nonsignificance of IgE glycosylation in receptor interaction. While existing literature underscores the significance of glycans at the N275 position for binding to FcεR1 receptor and initiation of anaphylaxis, the role of other IgE glycosylation sites in folding or receptor binding remains elusive. This study systematically investigates the functional significance of N-linked glycosylation sites in the heavy chain of IgE which validates the pivotal role of N275 residue in IgE secretion and stability. Replacement of this asparagine to non-amine group moieties does not affect IgE function in vitro, yet substitution with aspartic acid compromises antibody yield. The deglycosylated IgE variant exhibits superior efficacy, challenging the conventional importance of glycosylation for effector function. In summary, our study unveils an intricate relationship between N-glycosylation sites and the structural-functional dynamics of IgE antibodies. Furthermore, it offers novel insights into the IgE scaffold, paving the way for the development of more effective and stable IgE-based therapeutics.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":null,"pages":null},"PeriodicalIF":3.5,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142008334","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Glycosylation in Drosophila S2 cells. 果蝇 S2 细胞中的糖基化。

IF 3.5 2区生物学

Biotechnology and Bioengineering Pub Date : 2024-12-01 Epub Date: 2024-08-14 DOI: 10.1002/bit.28827

Tingting Xu, Lixiang Tong, Zhifu Zhang, Hairong Zhou, Peilin Zheng

{"title":"Glycosylation in Drosophila S2 cells.","authors":"Tingting Xu, Lixiang Tong, Zhifu Zhang, Hairong Zhou, Peilin Zheng","doi":"10.1002/bit.28827","DOIUrl":"10.1002/bit.28827","url":null,"abstract":"In recent years, there has been a remarkable surge in the approval of therapeutic protein drugs, particularly recombinant glycoproteins. Drosophila melanogaster S2 cells have become an appealing platform for the production of recombinant proteins due to their simplicity and low cost in cell culture. However, a significant limitation associated with using the S2 cell expression system is its propensity to introduce simple paucimannosidic glycosylation structures, which differs from that in the mammalian expression system. It is well established that the glycosylation patterns of glycoproteins have a profound impact on the physicochemical properties, bioactivity, and immunogenicity. Therefore, understanding the mechanisms behind these glycosylation modifications and implementing measures to address it has become a subject of considerable interest. This review aims to comprehensively summarize recent advancements in glycosylation modification in S2 cells, with a particular focus on comparing the glycosylation patterns among S2, other insect cells, and mammalian cells, as well as developing strategies for altering the glycosylation patterns of recombinant glycoproteins.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":null,"pages":null},"PeriodicalIF":3.5,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141975064","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Multidose transient transfection of human embryonic kidney 293 cells modulates recombinant adeno-associated virus2/5 Rep protein expression and influences the enrichment fraction of filled capsids. 多剂量瞬时转染人胚肾293细胞可调节重组腺相关病毒2/5 Rep蛋白的表达并影响填充囊壳的富集部分。

IF 3.5 2区生物学

Biotechnology and Bioengineering Pub Date : 2024-12-01 Epub Date: 2024-08-23 DOI: 10.1002/bit.28828

Prasanna Srinivasan, Christopher T Canova, Sha Sha, Tam N T Nguyen, John Joseph, Jose Sangerman, Andrew J Maloney, Georgios Katsikis, Rui Wen Ou, Moo Sun Hong, Jaclyn Ng, Arella Yuan, Daniel Antov, Sally Song, Wenyu Chen, Caleb Neufeld, Jacqueline M Wolfrum, Paul W Barone, Anthony J Sinskey, Stacy L Springs, Richard D Braatz

{"title":"Multidose transient transfection of human embryonic kidney 293 cells modulates recombinant adeno-associated virus2/5 Rep protein expression and influences the enrichment fraction of filled capsids.","authors":"Prasanna Srinivasan, Christopher T Canova, Sha Sha, Tam N T Nguyen, John Joseph, Jose Sangerman, Andrew J Maloney, Georgios Katsikis, Rui Wen Ou, Moo Sun Hong, Jaclyn Ng, Arella Yuan, Daniel Antov, Sally Song, Wenyu Chen, Caleb Neufeld, Jacqueline M Wolfrum, Paul W Barone, Anthony J Sinskey, Stacy L Springs, Richard D Braatz","doi":"10.1002/bit.28828","DOIUrl":"10.1002/bit.28828","url":null,"abstract":"Recombinant adeno-associated virus (rAAV) is a commonly used in vivo gene therapy vector because of its nonpathogenicity, long-term transgene expression, broad tropism, and ability to transduce both dividing and nondividing cells. However, rAAV vector production via transient transfection of mammalian cells typically yields a low fraction of filled-to-total capsids (~1%-30% of total capsids produced). Analysis of our previously developed mechanistic model for rAAV2/5 production attributed these low fill fractions to a poorly coordinated timeline between capsid synthesis and viral DNA replication and the repression of later phase capsid formation by Rep proteins. Here, we extend the model by quantifying the expression dynamics of total Rep proteins and their influence on the key steps of rAAV2/5 production using a multiple dosing transfection of human embryonic kidney 293 (HEK293) cells. We report that the availability of preformed empty capsids and viral DNA copies per cell are not limiting to the capsid-filling reaction. However, optimal expression of Rep proteins (<240 ± 13 ag per cell) enables enrichment of the filled capsid population (>12% of total capsids/cell) upstream. Our analysis suggests increased enrichment of filled capsids via regulating the expression of Rep proteins is possible but at the expense of per cell capsid titer in a triple plasmid transfection. Our study reveals an intrinsic limitation of scaling rAAV2/5 vector genome (vg) production and underscores the need for approaches that allow for regulating the expression of Rep proteins to maximize vg titer per cell upstream.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":null,"pages":null},"PeriodicalIF":3.5,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142035293","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Increase in polyunsaturated fatty acids and carotenoid accumulation in the microalga Golenkinia brevispicula (Chlorophyceae) by manipulating spectral irradiance and salinity. 通过调节光谱辐照度和盐度增加微藻 Golenkinia brevispicula（叶绿藻）中多不饱和脂肪酸和类胡萝卜素的积累。

IF 3.5 2区生物学

Biotechnology and Bioengineering Pub Date : 2024-12-01 Epub Date: 2024-08-25 DOI: 10.1002/bit.28831

T A Rearte, P S M Celis-Pla, R Abdala-Díaz, P Castro-Varela, S N Marsili, C García, M C Cerón-García, F L Figueroa

{"title":"Increase in polyunsaturated fatty acids and carotenoid accumulation in the microalga Golenkinia brevispicula (Chlorophyceae) by manipulating spectral irradiance and salinity.","authors":"T A Rearte, P S M Celis-Pla, R Abdala-Díaz, P Castro-Varela, S N Marsili, C García, M C Cerón-García, F L Figueroa","doi":"10.1002/bit.28831","DOIUrl":"10.1002/bit.28831","url":null,"abstract":"Microalgal biotechnology offers a promising platform for the sustainable production of diverse renewable bioactive compounds. The key distinction from other microbial bioprocesses lies in the critical role that light plays in cultures, as it serves as a source of environmental information to control metabolic processes. Therefore, we can use these criteria to design a bioprocess that aims to stimulate the accumulation of target molecules by controlling light exposure. We study the effect on biochemical and photobiological responses of Golenkinia brevispicula FAUBA-3 to the exposition of different spectral irradiances (specifically, high-fluence PAR of narrow yellow spectrum complemented with low intensity of monochromatic radiations of red, blue, and UV-A) under prestress and salinity stress conditions. High light (HL) intensity coupled to salinity stress affected the photosynthetic activity and photoprotection mechanisms as shown by maximal quantum yield (Fv/Fm) and non-photochemical quenching (NPQmax) reduction, respectively. HL treatments combined with the proper dose of UV-A radiation under salinity stress induced the highest carotenoid content (2.75 mg g dry weight [DW]- 1) composed mainly of lutein and β-carotene, and the highest lipid accumulation (35.3% DW) with the highest polyunsaturated fatty acid content (alpha-linolenic acid (C18:3) and linoleic acid (C18:2)). Our study can guide the strategies for commercial indoor production of G. brevispicula for high-value metabolites.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":null,"pages":null},"PeriodicalIF":3.5,"publicationDate":"2024-12-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142055003","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Biotechnology and Bioengineering: Volume 121, Number 12, December 2024 生物技术与生物工程第 121 卷第 12 号，2024 年 12 月

IF 3.8 2区生物学

Biotechnology and Bioengineering Pub Date : 2024-11-11 DOI: 10.1002/bit.28887

引用次数: 0

Correction to “A High‐Throughput Expression and Screening Platform for Applications‐Driven PETase Engineering” 对 "应用驱动 PETase 工程的高通量表达和筛选平台 "的更正

IF 3.8 2区生物学

Biotechnology and Bioengineering Pub Date : 2024-11-11 DOI: 10.1002/bit.28882

引用次数: 0

Mathematical Modeling and Simulation of 1,3‐Propanediol Production by Klebsiella pneumoniae BLh‐1 in a Batch Bioreactor Using Bayesian Statistics 利用贝叶斯统计对批式生物反应器中肺炎克雷伯氏菌 BLh-1 生产 1,3-丙二醇的过程进行数学建模和模拟

IF 3.8 2区生物学

Biotechnology and Bioengineering Pub Date : 2024-11-08 DOI: 10.1002/bit.28883

Nathalia Lobato Moraes, Mailson Batista de Vilhena, Daniele Misturini Rossi, Bruno Marques Viegas

{"title":"Mathematical Modeling and Simulation of 1,3‐Propanediol Production by Klebsiella pneumoniae BLh‐1 in a Batch Bioreactor Using Bayesian Statistics","authors":"Nathalia Lobato Moraes, Mailson Batista de Vilhena, Daniele Misturini Rossi, Bruno Marques Viegas","doi":"10.1002/bit.28883","DOIUrl":"https://doi.org/10.1002/bit.28883","url":null,"abstract":"Mathematical modeling and computer simulation are fundamental for optimizing biotechnological processes, enabling cost reduction and scalability, thereby driving advancements in the bioindustry. In this work, mathematical modeling and estimation of fermentative kinetic parameters were carried out to produce 1,3‐propanediol (1,3‐PDO) from residual glycerol and Klebsiella pneumoniae BLh‐1. The Markov chain Monte Carlo method, using the Metropolis‐Hastings algorithm, was applied to experimental data from a batch bioreactor under aerobic and anaerobic conditions. Sensitivity analysis and parameter evolution studies were conducted. The root‐mean‐square error (rRMSE) was chosen as the validation and calibration metric for the developed mathematical model. The results indicated that the average tolerance of glycerol was 174.68 and 44.85 g L−1, the inhibitory products was 150.95 g L−1 for ethanol and 35.56 g L−1 for 1,3‐PDO, and the maximum specific rate of cell growth was 0.189 and 0.275 h−1, for aerobic and anaerobic cultures, respectively. The model presented excellent fits in both crops, with rRMSE values between 0.09 − 33.74% and 3.58 − 31.82%, for the aerobic and anaerobic environment, respectively. With this, it was possible to evaluate and extract relevant information for a better understanding and control of the bioprocess.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":null,"pages":null},"PeriodicalIF":3.8,"publicationDate":"2024-11-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142597611","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Implemention of Innovative Process Analytical Technologies to Characterize Critical Quality Attributes of Co‐Formulated Monoclonal Antibody Products 采用创新工艺分析技术表征共配单克隆抗体产品的关键质量属性

IF 3.8 2区生物学

Biotechnology and Bioengineering Pub Date : 2024-11-08 DOI: 10.1002/bit.28881

Apurva Godbole, Lyufei Chen, Jay Desai, Smita Raghava, Richard Ruzanski, Bhumit Patel, Emmanuel Appiah‐Amponsah, Hanzhou Feng

{"title":"Implemention of Innovative Process Analytical Technologies to Characterize Critical Quality Attributes of Co‐Formulated Monoclonal Antibody Products","authors":"Apurva Godbole, Lyufei Chen, Jay Desai, Smita Raghava, Richard Ruzanski, Bhumit Patel, Emmanuel Appiah‐Amponsah, Hanzhou Feng","doi":"10.1002/bit.28881","DOIUrl":"https://doi.org/10.1002/bit.28881","url":null,"abstract":"Characterizing co‐formulated monoclonal antibodies (mAbs) poses significant challenges in the pharmaceutical industry. Due to the high structural similarity of the mAbs, traditional analytical methods, compounded by the lengthy method development process, hinder product development and manufacturing efficiency. There is increasing critical need in the pharmaceutical industry to streamline analytical approaches, minimizing time and resources, ensuring a rapid clinical entry and cost‐effective manufacturing. This study investigates the application of process analytical technologies (PAT) to address such challenges. Our investigation introduces two complementary technologies, on‐line ultra‐performance liquid chromatography (online UPLC) and multimode fluorescence spectroscopy (MMFS), as potential PAT tools tailored for characterizing critical quality attributes (CQA) in co‐formulated mAb products. Specifically, the CQAs under evaluation include the total protein concentration of the mAbs within the co‐formulation and the ratio of mAb A to mAb B. Online UPLC enables direct and automated measurement of the CQAs through physical separation, while MMFS determines them in a non‐destructive and more swift manner based on chemometric modeling. We demonstrate these technologies' comparable performance to conventional methods, alongside substantial benefits such as reduced analytical turnaround time and decreased laboratory efforts. Ultimately, integrating them as innovative PAT tools expedites the delivery of therapeutic solutions to patients and enhances manufacturing efficiency, aligning with the imperative for swift translation of scientific discoveries into clinical benefits.","PeriodicalId":9168,"journal":{"name":"Biotechnology and Bioengineering","volume":null,"pages":null},"PeriodicalIF":3.8,"publicationDate":"2024-11-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142597612","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0