Article
Biochemistry & Molecular Biology
A. Sesilja Aranko, Hideo Iwai
Summary: This study introduces an IIST system based on salt-inducible conditional protein splicing using the intein from Halorhabdus utahensis for purification of small peptides. It also describes a method for amidation and N-15 labeling of a single domain antibody using the same system.
Article
Engineering, Environmental
Sejong Choi, Yeeun Lee, Jeongin Hwang, Dahyun Chun, Heebeom Koo, Yan Lee
Summary: This study proposes a self-cleaving protein, pH inteinN150, as a pH-responsive linker for selective drug delivery. Unlike chemical linkers, protein linkers can be incorporated into protein pharmaceuticals during expression. By cleaving pH inteinN150 at weakly acidic pH, intracellular protein penetration can be turned on or off. Furthermore, the apoptosis-inducing activity of hTRAIL can be selectively activated by cleaving pH inteinN150 adjacent to ABD at weakly acidic pH. This new protein linker can potentially control protein-based drugs in response to pH variations around inflammatory or cancerous tissues.
CHEMICAL ENGINEERING JOURNAL
(2023)
Article
Biotechnology & Applied Microbiology
Guang Zeng, Yinzhen Zheng, Ya Xiang, Run Liu, Xiaofeng Yang, Zhanglin Lin
Summary: Our study successfully purified four proteins and peptides using a novel salt-inducible self-assembling peptide tag scheme, with yields ranging from 12 to 87 mg/L. The yield and purity (72-97%) were comparable to the classical His-tag method, but without using a His-tag.
MICROBIAL CELL FACTORIES
(2023)
Article
Biochemistry & Molecular Biology
Vlad-Stefan Raducanu, Daniela-Violeta Raducanu, Yujing Ouyang, Muhammad Tehseen, Masateru Takahashi, Samir M. Hamdan
Summary: TSGIT is a fusion-tag system composed of both N- and C-terminal composite fusion tags, which streamline the cloning, expression, and purification procedures to efficiently purify full-length proteins and prevent truncated forms.
Article
Pharmacology & Pharmacy
Lei Wang, Yu Qiao, Huifang Zong, Lei Han, Yong Ke, ZhiDi Pan, Jie Chen, Jun Lu, Jinyao Li, Tianlei Ying, Baohong Zhang, Jianwei Zhu
Summary: This study successfully generated a bispecific antibody targeting EpCAM and CD3, which has potential therapeutic application in colorectal cancer. In vitro experiments demonstrated that it could activate and recruit T cells, exhibiting potent cytotoxicity. In vivo experiments showed similar pharmacokinetic parameters and tumor growth inhibition.
FRONTIERS IN PHARMACOLOGY
(2022)
Article
Biotechnology & Applied Microbiology
Sandra Haas, Monika Desombre, Frank Kirschhoefer, Matthias C. Huber, Stefan M. Schiller, Juergen Hubbuch
Summary: This manuscript investigates the purification protocols for an elastin-like protein (ELP) and evaluates their scalability and performance. The study finds that immobilized metal ion affinity chromatography (IMAC) achieves the highest purity and lowest nucleic acid contamination, while salt-induced precipitation offers a fast and simple alternative for large-scale production. Additional inverse transition cycling (ITC) purification leads to the highest purity but requires expensive temperature-dependent centrifugation steps.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2022)
Article
Biochemistry & Molecular Biology
Michal Nemergut, Rostislav Skrabana, Martin Berta, Andreas Plueckthun, Erik Sedlak
Summary: Maltose binding protein (MBP) is historically used as an expression tag to enhance solubility of fused proteins, with affinity purification improved by a protein-protein interaction approach using immobilized Designed Ankyrin Repeat Protein off7 (DARPin off7). This new method simplifies the purification process of MBP fusion proteins, allowing for easy construction, resistance to amylase, insensitivity to maltose, and multiple reuse of the affinity matrix.
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
(2021)
Article
Engineering, Chemical
Zhanglin Lin, Yanyun Jing, Yuan Huang, Sha Yang, Shuyi Chen, Yingying Ou, Marco Pistolozzi, Xiaofeng Yang
Summary: This study provides a standardized method based on the cSAT scheme for the high-yield production of traditionally challenging disulfide bonded proteins and peptides. The method allows the proteins and peptides to correctly fold in an aggregated form during expression and enables high purity and high activity products through column purification.
CHEMICAL ENGINEERING SCIENCE
(2022)
Article
Biochemistry & Molecular Biology
Chih-Lung Wu, Ya-Han Chih, Hsin-Ying Hsieh, Kuang-Li Peng, Yi-Zong Lee, Bak-Sau Yip, Shih-Che Sue, Jya-Wei Cheng
Summary: Cecropins, a type of antimicrobial peptides found in the innate immune system of Cecropia moths, exhibit broad-spectrum antimicrobial and anticancer activities. This study developed a strategy to produce recombinant KR12AGPWR6 with enhanced antimicrobial properties through a self-cleavage system. The results showed that the recombinant KR12AGPWR6 exhibited similar antimicrobial activities compared to the chemically synthesized peptide. This method provides a potential strategy for large-scale production of AMPs.
Article
Biotechnology & Applied Microbiology
Sergio Enriquez-Flores, Jose Ignacio De la Mora-De la Mora, Luis Antonio Flores-Lopez, Nallely Cabrera, Cynthia Fernandez-Lainez, Gloria Hernandez-Alcantara, Carlos Enrique Guerrero-Beltran, Gabriel Lopez-Velazquez, Itzhel Garcia-Torres
Summary: This study reviewed the mutations made to the protease catalytic subunit of nuclear inclusion protein A from tobacco etch virus (TEVp) and created a mutant called TEVp7M. The mutant showed high purification yields and stability, and was able to cleave tagged proteins in commonly used affinity purification buffers.
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
(2022)
Article
Chemistry, Multidisciplinary
Baochang Zhang, Yupeng Zheng, Guochao Chu, Xiangyu Deng, Tongyue Wang, Weiwei Shi, Yongkang Zhou, Shan Tang, Ji-Shen Zheng, Lei Liu
Summary: This article presents a new method called Backbone-Installed Split Intein-Assisted Ligation (BISIAL) for the synthesis of membrane-associated D-proteins. The BISIAL method allows smooth ligation under strongly chaotropic conditions and the removal of ligation scars on the products, enabling the synthesis of challenging Dprotein targets.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2023)
Article
Chemistry, Multidisciplinary
Jan Pille, Antonio Aloi, Duc H. T. Le, Ilia Vialshin, Nathalie van de Laar, Kirsten Kevenaar, Maarten Merkx, Ilja K. Voets, Jan C. M. van Hest
Summary: This study introduces a novel pathway-dependent assembly method based on stimulus-responsive polymers, which can form monodisperse and stable polymeric particles under specific heating conditions. This method shows high efficiency in encapsulating cargo, such as fluorescent proteins and bioluminescent enzymes, and maintaining their function within cells.
Article
Chemistry, Multidisciplinary
Vusala Ibrahimova, Hang Zhao, Emmanuel Ibarboure, Elisabeth Garanger, Sebastien Lecommandoux
Summary: A new class of hybrid lipoprotein mimics was designed using chemical post-modification reactions, allowing the formation of biomimetic vesicles with controlled permeability, correlated to the temperature-responsiveness of ELPs.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2021)
Article
Biochemistry & Molecular Biology
Devin M. Ray, Julia R. Flood, Yael David
Summary: Biochemical studies of integral membrane proteins often face challenges of low purification yields and technical limitations. This study demonstrates the potential of using intein biorthogonality to modify target membrane proteins in live cells, bypassing these limitations and broadening the scope of accessible questions. The experiments using EGFR as a model show the efficiency and specificity of the split-intein pair Ava(N)/Npu(C) in modifying target membrane proteins for downstream live cell applications.
Review
Pharmacology & Pharmacy
Like Gong, Zhaoying Yang, Fan Zhang, Weiping Gao
Summary: Conjugation of cytokines with elastin-like polypeptides (ELPs) has shown great potential in addressing challenges such as poor stability, short circulation half-life, and difficulty in production. This approach offers extended circulation time, enhanced stability, increased soluble protein expression, simplified purification, improved drug delivery, and controlled release, with promising applications in the field.
ADVANCED DRUG DELIVERY REVIEWS
(2022)