Biotechnology Journal最新文献_第9页

A Split sfGFP-Based Signal-Amplified Reporter System for Enhanced Detection of Promoter Activity 基于分裂sfgfp的信号放大报告系统增强启动子活性检测。

IF 3.1 3区生物学

Biotechnology Journal Pub Date : 2026-05-19 DOI: 10.1002/biot.70244

Shuo Lin, Hui Yuan, Ziyi Lin, Shuangpeng Li, Yue Chi, Haihua Yin, Xia Zuo, Ruilin Cheng, Yuqian Tang, Zihao Luo, Min Chen, Yaoshun Liang, Xiaoqing Zhou, Chengcheng Tang, Qingjian Zou

{"title":"A Split sfGFP-Based Signal-Amplified Reporter System for Enhanced Detection of Promoter Activity","authors":"Shuo Lin, Hui Yuan, Ziyi Lin, Shuangpeng Li, Yue Chi, Haihua Yin, Xia Zuo, Ruilin Cheng, Yuqian Tang, Zihao Luo, Min Chen, Yaoshun Liang, Xiaoqing Zhou, Chengcheng Tang, Qingjian Zou","doi":"10.1002/biot.70244","DOIUrl":"10.1002/biot.70244","url":null,"abstract":"<div>\u0000 \u0000 <p>Fluorescent reporter systems for protein expression enable real-time, dynamic, and single-cell visualization of gene expression in living cells, but lack sufficient sensitivity for detecting low-abundance proteins. Herein, we developed a signal-amplified split superfolder green fluorescent protein (sfGFP) reporter system: sfGFP was split into the GFP<sub>1–10</sub> fragment driven by a constitutively strong promoter and a 20× tandem GFP<sub>11</sub> tag controlled by weak promoters, with the incorporation of coiled-coil dimerization domains to facilitate fragment complementation. This design enhanced fluorescent signals by approximately 10-fold, significantly improving detection sensitivity and enabling clear visualization of green fluorescence even at extremely low mRNA levels. Application of this system to evaluate Tet-On inducible systems revealed substantial basal leakage in Tet-On 2G. Moreover, real-time monitoring of endogenous weakly promoters (e.g., VIM, HPRT1, GUSB) demonstrated that the fluorescent intensity was approximately 10-fold higher than that of conventional reporter systems. This platform provides a highly sensitive and versatile tool for weak promoter activity analysis, dynamic tracking of low-abundance genes, and investigation of cell types and states.</p>\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"21 5","pages":""},"PeriodicalIF":3.1,"publicationDate":"2026-05-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147969074","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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

Harnessing Nature's Algorithm: From Test Tubes to Autonomous In Vivo Evolution 利用自然算法：从试管到自主体内进化。

IF 3.1 3区生物学

Biotechnology Journal Pub Date : 2026-05-19 DOI: 10.1002/biot.70237

Wenna Shang, Zhengbing Lyu, Guodong Chen

{"title":"Harnessing Nature's Algorithm: From Test Tubes to Autonomous In Vivo Evolution","authors":"Wenna Shang, Zhengbing Lyu, Guodong Chen","doi":"10.1002/biot.70237","DOIUrl":"10.1002/biot.70237","url":null,"abstract":"<div>\u0000 \u0000 <p>Directed evolution (DE) enables the engineering of biomolecules without prior structural knowledge. However, traditional step-wise DE is constrained by limited screening throughput. To more efficiently navigate epistatic fitness landscapes, the field is increasingly adopting autonomous, continuous in vivo evolution systems. This review critically examines the molecular architectures and engineering principles driving this transition. We evaluate strategies for continuous genetic diversification—ranging from orthogonal replication systems (e.g., OrthoRep, T7-ORACLE) to CRISPR-guided mutagenesis (e.g., EvolvR)—with a focus on the fundamental trade-off between mutational load and host viability. Furthermore, we analyze the biophysical constraints of screening and the kinetic demands of coupling real-time selection with ultra-fast mutagenesis, as exemplified by phage-assisted continuous evolution (PACE). Crucially, we explore the functional integration of machine learning (ML), highlighting how active learning models and zero-shot predictions via protein language models (PLMs) can resolve epistatic complexities and mitigate the latency of next-generation sequencing. Finally, we discuss the multidimensional hardware and algorithmic bottlenecks currently impeding the realization of fully closed-loop biofoundries, and assess the strategic implications of these technologies for accelerating the engineering of complex therapeutics.</p>\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"21 5","pages":""},"PeriodicalIF":3.1,"publicationDate":"2026-05-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147969094","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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

IF 3.1 3区生物学

Biotechnology Journal Pub Date : 2026-05-19

引用次数: 0

IF 3.1 3区生物学

Biotechnology Journal Pub Date : 2026-05-19

引用次数: 0

Oral Lactobacillus reuteri Combined With Intravenous Lysis-Circuit-Bearing Engineered Bacteria RE Enhances Antitumor Efficacy 口服罗伊氏乳杆菌联合静脉溶袢工程菌RE增强抗肿瘤效果。

IF 3.1 3区生物学

Biotechnology Journal Pub Date : 2026-05-19 DOI: 10.1002/biot.70243

Feng Geng, Lijun Yue, Maoru Du, Jiayi Jin, Qiongwen Zhang, Wenxin Hu, Na Hao, Sitong Liu, Peilin Jiang, Xiaohong Pan

{"title":"Oral Lactobacillus reuteri Combined With Intravenous Lysis-Circuit-Bearing Engineered Bacteria RE Enhances Antitumor Efficacy","authors":"Feng Geng, Lijun Yue, Maoru Du, Jiayi Jin, Qiongwen Zhang, Wenxin Hu, Na Hao, Sitong Liu, Peilin Jiang, Xiaohong Pan","doi":"10.1002/biot.70243","DOIUrl":"10.1002/biot.70243","url":null,"abstract":"<div>\u0000 \u0000 <p>Engineered bacteria can preferentially colonize hypoxic tumors, but improving efficacy while limiting systemic bacterial burden remains a central challenge. Here, we combined oral <i>Lactobacillus reuteri</i> (LR) with intravenous administration of RE, an engineered <i>Escherichia coli Rosetta</i> strain carrying the quorum-sensing lysis plasmid pTD103-E. This circuit couples LuxI/R-mediated population sensing to expression of the phage lysis gene φX174 E and was designed to enable density-associated bacterial lysis. In a 4T1 mouse model, LR+RE improved antitumor efficacy by 45% compared with single-bacterium treatment. RE remained enriched in tumors while being cleared from major organs within 72 h. Combined treatment increased intratumoral IFN-γ, altered IL-12 expression, elevated AhR abundance in tumor tissues, and increased CD8+ T-cell infiltration. These changes were accompanied by polarization of tumor-associated macrophages toward an M1-like phenotype, suppression of M2-like features, and increased markers of cell-cycle arrest and apoptosis. Fecal 16S rRNA profiling further showed that treatment reshaped the gut microbiota and identified <i>Prevotellaceae_NK3B31_group</i> as a microbial taxon associated with IFN-γ-centered antitumor responses. Together, these findings indicate that a natural probiotic can potentiate a tumor-targeted engineered bacterial therapy and is associated with coordinated microbiota-immune remodeling.</p>\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"21 5","pages":""},"PeriodicalIF":3.1,"publicationDate":"2026-05-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147969017","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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

IF 3.1 3区生物学

Biotechnology Journal Pub Date : 2026-05-19

引用次数: 0

IF 3.1 3区生物学

Biotechnology Journal Pub Date : 2026-05-13

引用次数: 0

Establishment and Comparison of High-Throughput Immunoassays for Quantifying Clostridioides difficile Infection Biomarkers 艰难梭菌感染生物标志物高通量免疫测定方法的建立与比较。

IF 3.1 3区生物学

Biotechnology Journal Pub Date : 2026-05-13 DOI: 10.1002/biot.70238

Hanool Yun, Na-Hyeon Kwon, Minhee Kang, Tae Yeul Kim, Hee-Jin Jeong

{"title":"Establishment and Comparison of High-Throughput Immunoassays for Quantifying Clostridioides difficile Infection Biomarkers","authors":"Hanool Yun, Na-Hyeon Kwon, Minhee Kang, Tae Yeul Kim, Hee-Jin Jeong","doi":"10.1002/biot.70238","DOIUrl":"10.1002/biot.70238","url":null,"abstract":"<div>\u0000 \u0000 <p><i>Clostridioides difficile</i> infection (CDI) is the leading cause of healthcare-associated diarrhea, most often occurring after antibiotic exposure. In this study, we established three immunoassays: indirect enzyme-linked immunosorbent assay (ELISA), sandwich ELISA, and fluorescence-linked immunosorbent assay (FLISA), using antibodies specific to the major virulence factors, toxin A (TcdA) and toxin B (TcdB). Indirect ELISA demonstrated high specificity with an overall assay time within 21 h, achieving sensitive detection of both toxins. Although more time- and resource-intensive (22 h), sandwich ELISA is advantageous for applications where pre-coated plates or ready-to-use membranes can reduce assay time. FLISA, using optimized fluorescent dye-conjugated antibodies, offered the most rapid workflow and simplest preparation, with potential for multiplex detection, though its sensitivity was lower than that of ELISA. All three assays were implemented in a 96-well plate format, which supported parallel sample processing, reduced turnaround times, and offered scalability for high-throughput applications. Depending on assay requirements, such as analytical sensitivity, turnaround time, and cost-effectiveness, these methods provide complementary approaches for the detection of TcdA and TcdB.</p>\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"21 5","pages":""},"PeriodicalIF":3.1,"publicationDate":"2026-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147924598","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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

IF 3.1 3区生物学

Biotechnology Journal Pub Date : 2026-05-13

引用次数: 0

Regression Models of Viability and Apoptosis Can be Generated Leveraging Cluster Analysis From Ovizio iLineF PRO Image Data 利用Ovizio iLineF PRO图像数据的聚类分析可以生成细胞活力和细胞凋亡的回归模型。

IF 3.1 3区生物学

Biotechnology Journal Pub Date : 2026-05-13 DOI: 10.1002/biot.70231

Joseph Fantuzzo, Nicholas Zill, John Bowers, Balrina Gupta

{"title":"Regression Models of Viability and Apoptosis Can be Generated Leveraging Cluster Analysis From Ovizio iLineF PRO Image Data","authors":"Joseph Fantuzzo, Nicholas Zill, John Bowers, Balrina Gupta","doi":"10.1002/biot.70231","DOIUrl":"10.1002/biot.70231","url":null,"abstract":"<div>\u0000 \u0000 <p>Chinese hamster ovary (CHO) cells serve as the backbone of modern large-scale manufacturing of monoclonal antibodies. Central to this process are fed-batch cultures, where cells are grown from low to high cell densities and go through a mAb production phase. Despite CHO cells' widespread usage and vital role in the production of biologics, the cellular states during fed-batch coinciding with high specific productivity and apoptosis are poorly understood. Crucial to this understanding is a clear depiction of the various subpopulations of cells that exist in a fed-batch culture over time. In this work, an Ovizio iLine F PRO was used to image the cells in benchtop bioreactors and gather morphological and optical information for several CHO cell lines. A cluster analysis was applied to the Ovizio iLine F PRO raw data, revealing a diverse set of cellular sub-populations within each culture. Raw data from the Ovizio were then transformed into tank-level data and applied to offline measurements for viability and apoptosis in regression analysis. We used the results from the cluster analysis and feature averaging to set up regressions for offline measurements. Our regression analysis illustrates the power of in-line imaging of CHO cells as a rich process analytical tool.</p>\u0000 </div>","PeriodicalId":134,"journal":{"name":"Biotechnology Journal","volume":"21 5","pages":""},"PeriodicalIF":3.1,"publicationDate":"2026-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147924684","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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