Article
Biotechnology & Applied Microbiology
Amruta A. Shendge, Jacinta S. D'Souza
Summary: Expressing affinity-tagged recombinant proteins at a high yield is crucial for various applications. Escherichia coli remains the most popular expression system due to its advantages in growth, genetic manipulation, and cost. However, some proteins tend to form inclusion bodies when over-expressed in E. coli, leading to purification and yield challenges. This study focuses on solubilizing hydrophobic peptides and proteins with amphipathic helices using a combinatorial approach of chemical and physical methods.
MICROBIAL CELL FACTORIES
(2022)
Article
Biochemical Research Methods
Adria Lopez-Cano, Paula Sicilia, Clara Gaja, Anna Aris, Elena Garcia-Fruitos
Summary: Recombinant protein production in bacteria often leads to the formation of inclusion bodies (IBs). While strategies to minimize aggregation have been developed, many heterologous proteins still form aggregates. The detergent NLS is effective in solubilizing proteins from IBs, but its impact on protein quality has not been evaluated. This study compared the activity of antimicrobial proteins solubilized with NLS from IBs to those solubilized without detergent, and found that protein activity was impaired by NLS. The results suggest that the solubilization protocol used for IBs can impact the final protein quality.
Article
Biotechnology & Applied Microbiology
G. C. Lima, R. M. Chura-Chambi, L. Morganti, V. J. Silva, M. P. Cabral-Piccin, V. Rocha, T. S. Medina, R. N. Ramos, D. Luz
Summary: This study successfully overcame the challenge of recovering functional proteins from bacterial inclusion bodies and produced an active, non-glycosylated TIM-3 ectodomain. This protein can be used to investigate the interactions and functions of immune checkpoints.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2023)
Review
Biotechnology & Applied Microbiology
Arshpreet Bhatwa, Weijun Wang, Yousef I. Hassan, Nadine Abraham, Xiu-Zhen Li, Ting Zhou
Summary: Recombinant proteins are crucial in industrial applications, with E. coli being the primary bacterial host for their production. The challenge of inclusion body formation can be overcome through various strategies, such as adjusting growth conditions and modifying proteins of interest. Understanding the unique features and formation mechanism of inclusion bodies will help maximize their potential advantages.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2021)
Review
Biotechnology & Applied Microbiology
Yin Yin Siew, Wei Zhang
Summary: The Global Diabetes Compact initiated by the World Health Organization aims to improve accessibility and affordability of life-saving insulin. As the demand for recombinant insulin therapeutics rises globally, the downstream processing from E. coli inclusion bodies plays a crucial role in production efficiency. A comprehensive review of the downstream processing of recombinant human insulin/analogue production has been presented, highlighting the importance of optimizing purification schemes for more efficient and cost-effective insulin production.
BIORESOURCES AND BIOPROCESSING
(2021)
Article
Biochemistry & Molecular Biology
Jose Vicente Carratala, Laia Gifre-Renom, Ramon Roca-Pinilla, Antonio Villaverde, Anna Aris, Elena Garcia-Fruitos, Julieta Maria Sanchez, Neus Ferrer-Miralles
Summary: The study analyzed the physicochemical characteristics of MMP-9 protein species, identifying four protein peaks and using DLS, CD, and spectrofluorometric analysis to confirm the separation of conformers with specific characteristics. This approach provides valuable information for selecting the most suitable protein populations based on their individual physiochemical properties and biological activity.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2021)
Article
Biology
Victor Marchenkov, Elena Dubovitskya, Nina Kotova, Igor Tuchkov, Nina Smirnova, Natalia Marchenko, Alexey Surin, Vladimir Filimonov, Gennady Semisotnov
Summary: The study proposes the use of transverse urea gradient electrophoresis to test the folding capability of recombinant proteins from inclusion bodies dissolved in urea. A successful refolding protocol was developed based on this method, providing a new approach for large-scale protein recovery from inclusion bodies.
Review
Biotechnology & Applied Microbiology
Suraj Mital, Graham Christie, Duygu Dikicioglu
Summary: Recombinant enzyme expression in Escherichia coli is a popular method for producing protein products, but is often limited by inclusion body formation. This study reviews literature from 2010 to 2021 and highlights the lack of a coherent strategy for promoting solubility in DtE enzymes, suggesting a need for a systematic approach using modern bioinformatics and 'omics' techniques. The analysis also points out potential gaps in reporting metadata in publications, which could impact reproducibility and growth in this field.
MICROBIAL CELL FACTORIES
(2021)
Article
Biotechnology & Applied Microbiology
Qi Wang, Chun Zhang, Zenglan Li, Fangxia Guo, Jing Zhang, Yongdong Liu, Zhiguo Su
Summary: The study successfully refolded the highly hydrophobic protein rhIFN beta-lb IBs using a strategy of high hydrostatic pressure combined with additives, achieving a good yield. The resulting product showed similar structures and in vitro bioactivity to the standard product.
BIOCHEMICAL ENGINEERING JOURNAL
(2021)
Article
Biochemistry & Molecular Biology
Dilek Sahinbas, Eda Celik
Summary: In this study, IL-6 specific single-chain antibodies were successfully expressed in E. coli, and the purity and activity of the product were improved through an optimized method. The results of this study are of significant importance for the diagnosis and treatment of inflammatory diseases.
PROCESS BIOCHEMISTRY
(2023)
Article
Biochemical Research Methods
Kajal Kachhawaha, Santanu Singh, Khyati Joshi, Priyanka Nain, Sumit K. Singh
Summary: This review provides a comprehensive overview of the bio-processing of bacterial inclusion bodies, including potential solutions to address critical issues. The improved understanding of the structure-function relationship of the inclusion bodies has led to the development of novel biotechnologies, promising to revolutionize therapeutic protein production.
PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY
(2023)
Article
Biochemical Research Methods
Brianne E. Bell, Isaac K. Burton, Jose Arreola-Patino, Thomas I. Harris, Paula Oliveira, Dong Chen, Randolph V. Lewis, Justin A. Jones
Summary: This study successfully developed a scalable method to purify recombinant hagfish intermediate filament proteins with increased protein recovery and purity, without interfering with fiber production or formation.
PROTEIN EXPRESSION AND PURIFICATION
(2022)
Article
Microbiology
Nguyen Thi My Trinh, Tran Linh Thuoc, Dang Thi Phuong Thao
Summary: Producing G-CSF in E. coli as non-classical inclusion bodies (ncIBs) has been shown to be a feasible approach to improve yield and lower manufacturing costs, providing a protein with high purity and similar bioactivity to commercially available products like Neupogen.
BRAZILIAN JOURNAL OF MICROBIOLOGY
(2021)
Article
Biochemical Research Methods
Zachary J. Knepp, Ashlea Ghaner, Kyle T. Root
Summary: This study presents a simple strategy for isolating cold-active recombinant esterase enzyme when expressed as inclusion bodies. The refolded AaSGNH1 enzyme was characterized, showing similar kinetic properties to other SGNH esterases, confirming its effectiveness.
PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY
(2022)
Article
Biochemical Research Methods
Guang Li, Weiping Li, Xiaolan Fang, Xuri Song, Shujing Teng, Zong Ren, Daoqi Hu, Songhui Zhou, Gangqiang Wu, Keqiang Li
Summary: Antibody detection in COVID-19 diagnosis offers advantages of convenience, high throughput, and low cost when combined with nucleic acid testing. Virus infection stimulates high antibody titer against SARS-CoV-2 nucleocapsid protein, which can be used to detect COVID-19 in both infected and convalescent patients.
PROTEIN EXPRESSION AND PURIFICATION
(2021)
Article
Biochemical Research Methods
Juri Sakata, Toshifumi Tatsumi, Akira Sugiyama, Akihiro Shimizu, Yuya Inagaki, Hiroto Katoh, Takefumi Yamashita, Kazuki Takahashi, Sho Aki, Yudai Kaneko, Takeshi Kawamura, Mai Miura, Masazumi Ishii, Tsuyoshi Osawa, Toshiya Tanaka, Shumpei Ishikawa, Masanobu Tsukagoshi, Michael Chansler, Tatsuhiko Kodama, Motomu Kanai, Hidetoshi Tokuyama, Kenzo Yamatsugu
Summary: This study reports an expression, refolding, and purification method for antibody-mimetic drug conjugate (AMDC). The AMDC uses a variable heavy chain of heavy chain-only antibodies (VHHs) as the targeting protein and non-covalently binds to a potent DNA-alkylating drug. The AMDC shows strong cytotoxic effects on cancer cells.
PROTEIN EXPRESSION AND PURIFICATION
(2024)
Article
Biochemical Research Methods
Yao Wang, Shaoting Weng, Yajie Tang, Sen Lin, Xiayue Liu, Wenhui Zhang, Gang Liu, Boomi Pandi, Yinrong Wu, Lei Ma, Lin Wang
Summary: In this study, a high yield of recombinant CD20 and claudin 18.2 proteins was achieved using an in vitro coupled transcription-translation system. The results showed that rituximab has a high affinity with CD20 protein. This study provides a novel concept for promoting the expression of multi-pass transmembrane proteins and lays the foundation for large-scale industrial production of membrane-associated drug targets.
PROTEIN EXPRESSION AND PURIFICATION
(2024)
Article
Biochemical Research Methods
Xiaobing Chen, Zijuan Zou, Wei Li, Xu Dong, Yi Chen, Yan Lu, Mingyue Zhu, Mengsen Li, Bo Lin
Summary: In this study, the fusion protein ImI-AFP3, composed of alpha-Conotoxin ImI and human alpha fetoprotein domain 3 (AFP3), was found to inhibit the growth and migration of lung cancer cells and showed synergistic effects with the drug gefitinib. These findings suggest that ImI-AFP3 is a promising candidate for the development of anticancer drugs.
PROTEIN EXPRESSION AND PURIFICATION
(2024)
Article
Biochemical Research Methods
Angel Castillo-Corujo, Mirva J. Saaranen, Lloyd W. Ruddock
Summary: This study successfully expressed two Fabs antibodies in the cytoplasm of E. coli using the CyDisCo system, achieving high yields and biological activity under industrially relevant fermentation conditions.
PROTEIN EXPRESSION AND PURIFICATION
(2024)
Article
Biochemical Research Methods
Junling Guo, Zhongyi Cheng, Zhemin Zhou
Summary: This study focused on a halophilic archaeal nitrile hydratase (NHase) and found that it exhibited higher tolerance to substrates and products compared to NHases from other sources. The unique genetic structure of this highly stable archaeal NHase could provide a theoretical foundation for modifying and enhancing the industrial application of NHase.
PROTEIN EXPRESSION AND PURIFICATION
(2024)
Article
Biochemical Research Methods
Yunlong Shen, Ruirui Zhang, Xiaohua Jiang, Jinliang Yang
Summary: The study developed a monoclonal antibody, B1M023, that can bind to LILRB1 with high affinity and block its binding to HLA-G. This antibody can promote the activation and IFN-gamma secretion of T cells, suggesting its potential applications in concomitant diagnosis and tumor immunotherapy.
PROTEIN EXPRESSION AND PURIFICATION
(2024)
Article
Biochemical Research Methods
Jun Jiang, Zhengqiang Jiang, Qiaojuan Yan, Susu Han, Shaoqing Yang
Summary: A novel alginate lyase gene from a marine bacterium showed efficient enzyme activity and stability, producing high conversion ratios of alginate oligosaccharides.
PROTEIN EXPRESSION AND PURIFICATION
(2024)
Article
Biochemical Research Methods
Xianghua Xiong, Yujin Qiu, Jiahao Zheng, Ling Zhou, Qingyang Wang, Jinglun Pang, Weicai Zhang, Huipeng Chen, Gang Liu, Xiaodong Han
Summary: A specific monoclonal antibody ML419 has been found to disrupt the recognition between Botulinum neurotoxin serotype A (BoNT/A) and FGFR3, effectively preventing BoNT/A from entering neurons. In vivo experiments show that ML419 has a strong protective effect, making it a promising candidate for the development of therapeutics against BoNT/A.
PROTEIN EXPRESSION AND PURIFICATION
(2024)
Article
Biochemical Research Methods
Maria Rain Jennings, Soohyon Min, Grace S. Xu, Kassandra Homayuni, Bhavana Suresh, Yusef Amir Haikal, John Blazeck
Summary: This study successfully produced Homo sapiens adenosine deaminase isoform 1 (HsADA1) and its variants through optimizing the recombinant expression process. The D8N variant of HsADA1 was found to be about 30% less active than the wildtype, but it better retained its activity in human serum. Additionally, the study revealed a previously undescribed phenomenon involving albumin that contributed to the increased activity of HsADA1 and the D8N variant in serum.
PROTEIN EXPRESSION AND PURIFICATION
(2024)
Article
Biochemical Research Methods
Oinam Sangita Devi, Senjam Sunil Singh, K. Rana, Sorokhaibam Jibankumar Singh, Wayenbam Sobhachandra Singh
Summary: A new lectin with hemagglutination activity was purified from the rhizome of Xanthosoma violaceum Schott. The lectin showed different reactions towards human red blood cells of different blood groups. It exhibited optimal hemagglutination activity at specific temperature and pH range, and showed good stability.
PROTEIN EXPRESSION AND PURIFICATION
(2024)