Article
Biochemical Research Methods
Yoko Ino, Yutaro Yamaoka, Kiho Tanaka, Kei Miyakawa, Mayuko Nishi, Yasuyoshi Hatayama, Hirokazu Kimura, Yayoi Kimura, Akihide Ryo
Summary: Peptide tag systems are widely used for protein detection and purification. We developed a novel tag system called HiP4 with a specific epitope sequence that overlaps with the conventional 6x histidine tag. By using this system, we developed a TAP-MS system for comprehensive protein interactome analysis, which showed low background and high selectivity in identifying novel protein interactions.
Article
Neurosciences
Jia-Hua Hu, Ying Liu, Dax A. Hoffman
Summary: Proteins in neurons form complexes to regulate signal transduction and synaptic function. A novel method combining lentiviral protein expression with tandem affinity purification followed by mass-spectrometry (TAP-MS) has been successfully used to identify protein complexes and post-translational modifications in neurons, shedding light on potential mechanisms underlying neurological diseases.
FRONTIERS IN CELLULAR NEUROSCIENCE
(2022)
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
Multidisciplinary Sciences
Susan K. Vester, Rolle Rahikainen, Irsyad N. A. Khairil Anuar, Rory A. Hills, Tiong Kit Tan, Mark Howarth
Summary: The authors developed a switchable protein purification method called SpySwitch, which enables gentle purification of proteins tagged with SpyTag. SpySwitch contains histidines that allow pH-dependent release of captured proteins, resulting in higher purity compared to traditional His-tag methods. Additionally, SpySwitch is thermosensitive, allowing protein capture and release at specific temperatures, making it suitable for direct assembly onto multimeric scaffolds. The study also demonstrated the application of SpySwitch in the purification of receptor-binding domains from coronaviruses and the recognition of these domains by antibodies.
NATURE COMMUNICATIONS
(2022)
Article
Biochemistry & Molecular Biology
Attila Meszaros, Kevin Muwonge, Steven Janvier, Junaid Ahmed, Peter Tompa
Summary: Intrinsically disordered proteins (IDPs) lack well-defined 3D structures and can interact promiscuously due to their high flexibility. This unique property allows them to play diverse regulatory roles in cellular processes. However, their exposed structural state makes them more susceptible to proteolytic degradation compared to globular proteins.
Article
Biochemical Research Methods
Guizhen Liu, Yanan Du, Tao Fu, Ying Han, Lifeng Pan, Jingwu Kang
Summary: The study presented a method for profiling protein-protein interactions using cm-IMAC columns in combination with label-free quantitative proteomics, successfully exploring protein-protein interactions and identifying new interactors of Bcl-X L.
JOURNAL OF CHROMATOGRAPHY A
(2022)
Article
Nanoscience & Nanotechnology
Yasmin Kaveh-Baghbaderani, Raphaela Allgayer, Sebastian Patrick Schwaminger, Paula Fraga-Garcia, Sonja Berensmeier
Summary: The study introduces a nanoparticle-based material for efficient magnetic separation of IgG antibodies with high binding capacity. Utilizing optimized protein A-based ligands directly immobilized on cost-effective bare iron oxide nanoparticles, the material allows for selective and high-recovery antibody capture, surpassing current technologies in both recovery rate and cost-effectiveness.
ACS APPLIED NANO MATERIALS
(2021)
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
Biotechnology & Applied Microbiology
Xiaomei He, Shuncheng Zhang, Dongya Dang, Tingting Lin, Yuanyuan Ge, Xiaofeng Chen, Jun Fan
Summary: This study optimized the separation conditions for the hanA1-tagged EmGFP protein and successfully used five different colored proteins to detect the effectiveness of the hanA1 fusion handle. The results suggest that the fusion hanA1 tag could be potentially used for simple and cheap affinity separation of target proteins in industry and diagnosis.
MICROBIAL CELL FACTORIES
(2023)
Article
Engineering, Chemical
Ana Freitas, Lucilia Domingues, Tatiana Q. Aguiar
Summary: This study evaluated the feasibility of using bare silica as a matrix for protein purification/immobilization, and the results showed that unmodified silica matrices can effectively purify His-tagged proteins, with the combination of tags being advantageous for immobilization purposes.
SEPARATION AND PURIFICATION TECHNOLOGY
(2022)
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)
Review
Biochemical Research Methods
Rebecca Elizabeth Kattan, Deena Ayesh, Wenqi Wang
Summary: During intracellular signal transduction, protein-protein interactions (PPIs) play a critical role in regulating protein localization and function. Mass spectrometry-based interactome analysis has become a popular method for studying PPI networks, but the analysis of large datasets can be challenging. In this review, we discuss the methods and resources commonly used for analyzing large interactome-related proteomic data and propose a guideline for identifying novel interacting proteins.
BRIEFINGS IN BIOINFORMATICS
(2023)
Article
Biotechnology & Applied Microbiology
Stefan Schwarz, Doreen Gerlach, Rong Fan, Peter Czermak
Summary: This study tested the potential of Vibrio natriegens for producing antimicrobial peptides and found that it has advantages as an expression platform and for protein secretion and purification. Modification of the glucosamine-binding protein A tag increased the yield of antimicrobial peptides.
ELECTRONIC JOURNAL OF BIOTECHNOLOGY
(2022)
Article
Chemistry, Analytical
Jianan Feng, Linlin Jiang, Yiqing Cao, Chunhui Deng, Yan Li
Summary: In this study, a tractable method was established for rapid quality assessment of a harvested cell culture fluid (HCCF) sample by differentially extracting IgG with different Fc gamma RIIIa affinity levels and determining the amount and monomer percentage of IgGs using size exclusion chromatography (SEC).
ANALYTICAL CHEMISTRY
(2022)
Article
Chemistry, Multidisciplinary
Samuel L. Scinto, Tyler R. Reagle, Joseph M. Fox
Summary: This study demonstrates a new method for site-selective functionalization of proteins using pyridyl-tetrazine tags, allowing for direct affinity purification on commonly used nickel-iminodiacetate (Ni-IDA) resins. The method is also applicable for protein purification from complex mixtures.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2022)