Article
Chemistry, Multidisciplinary
Jonas Schilz, Charlotte Clement, Franziska Greiner, Arne Skerra
Summary: A novel strategy for the efficient and gentle affinity purification of PASylated proteins is reported, which allows direct one-step purification from cell extracts or culture supernatants while avoiding harsh elution conditions. This method can improve the efficiency of biological drug development.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2022)
Article
Chemistry, Analytical
Yu-ki Tanaka, Shunsuke Shimazaki, Yasunori Fukumoto, Yasumitsu Ogra
Summary: A rapid and precise quantification method for a histidine-tagged recombinant protein produced in Escherichia coli (E. coli) has been developed using single-cell inductively coupled plasma-mass spectrometry (SC-ICP-MS). This method allows for the determination of protein quantity based on the signal of nickel or cobalt ions bound to the His-tag, without the need for extraction and purification processes.
ANALYTICAL CHEMISTRY
(2022)
Article
Biochemical Research Methods
Merve Asena Ozbek, Erdogan Ozgur, Nilay Bereli, Adil Denizli
Summary: The demand for low-cost, easy, and efficient protein purification methods has been increasing in the field of protein purification, which is also important for enzyme production. An alternative approach using thrombin imprinted microcryogels has been developed for the purification of thrombin, allowing for reuse and high selectivity. Characterization studies were conducted using various techniques, and the maximum thrombin adsorption capacity was found to be 55.86 mg/g. The selectivity of the microcryogels was investigated, and the purity of thrombin was evaluated using Fast Performance Liquid Chromatography (FPLC).
JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES
(2023)
Article
Engineering, Chemical
M. J. B. Matos, F. Trovao, J. Goncalves, U. Rothbauer, M. G. Freire, A. M. J. B. Barbosa, A. S. Pina, A. C. A. Roque
Summary: Antibody technologies are a successful case in the biopharmaceutical industry, with researchers developing affinity adsorbents inspired by natural and engineered antibody-binding ligands to capture full-length antibodies and fragments in a single-step recovery process.
SEPARATION AND PURIFICATION TECHNOLOGY
(2021)
Article
Biotechnology & Applied Microbiology
Matthew J. McNulty, Naomi Hamada, Jesse Delzio, Liber McKee, Somen Nandi, Marjorie L. Longo, Karen A. McDonald
Summary: Advancements in understanding and engineering of virus-based nanomaterials (VBNs) for biomedical applications have led to the exploration of interfaces between VBNs and other biomedically-relevant chemistries and materials. The immobilization of antibody-binding VBN in silica sol-gel has shown promising results for bioseparations and biosensing applications.
JOURNAL OF NANOBIOTECHNOLOGY
(2022)
Article
Microbiology
Haiqiao Bian, Chong Yu, Yanwu Wei, Li Feng, Changming Liu, Liping Huang
Summary: The study successfully purified PCV2 virus strains using affinity chromatography, resulting in high purity and no significant changes in antigenic characteristics after purification. It was found that PCV2 viruses can be successfully purified, but the yield and purity may vary among different subtypes, and the purified virus can be used for virus structure analysis and viral adsorption and invasion research.
Article
Biochemistry & Molecular Biology
Saman Rahmati, Kowsar Bagherzadeh, Seyed Shahriar Arab, Fatemeh Torkashvand, Massoud Amanlou, Behrouz Vaziri
Summary: In this study, molecular dynamics simulations and computational biology methods were used to design high-affinity mutants of Protein L single B domains, which were then polymerized into a ligand with six B domains. The results showed that these mutants exhibited higher binding affinity to Fab compared to the wild type.
JOURNAL OF BIOMOLECULAR STRUCTURE & DYNAMICS
(2023)
Article
Biochemical Research Methods
Julia Scheffel, Sophia Hober
Summary: The manufacturability of therapeutic monoclonal antibodies is limited by harsh purification conditions, but a novel mild purification strategy using a calcium-dependent ligand derived from Protein A has been developed. By capturing antibodies with the tetrameric Z(Ca) resin and eluting with a chelator-free method, such as citrate, the elution process is significantly improved. Further optimization with sodium chloride can enhance elution efficiency, providing a cost-efficient and functional way to release antibodies.
JOURNAL OF CHROMATOGRAPHY A
(2021)
Article
Biochemical Research Methods
Zongqing Huang, Haoju Hua, Xiuzhen Du, Zipeng Zhen, Wei Zhao, Jun Feng, Ji-an Li
Summary: This study successfully isolated VHHs against the SUMO protein for the first time using biopanning of an immune camelid nanobody library. The VHHs were coupled to agarose resins to form affinity chromatography resins, allowing for one-step purification of various SUMO fusions.
JOURNAL OF CHROMATOGRAPHY A
(2024)
Article
Biophysics
Atsushi Hirano, Shogo Kanoh, Kentaro Shiraki, Momoyo Wada, Masahiro Kitamura, Katsuya Kato
Summary: This study discovered that phosphate-modified zirconia nanoparticles can selectively adsorb immunoglobulin G (IgG) antibodies with high capacity while maintaining their molecular conformation. Compared to traditional Protein A ligands, zirconia nanoparticles have higher affinity for human IgG antibodies, especially IgG3. Due to their chemical and mechanical stability, zirconia nanoparticles can be used in batch methods and chromatography as an alternative to Protein A chromatography for the purification of antibody drugs.
COLLOIDS AND SURFACES B-BIOINTERFACES
(2023)
Article
Biotechnology & Applied Microbiology
Matthew J. McNulty, Anton Schwartz, Jesse Delzio, Kalimuthu Karuppanan, Aaron Jacobson, Olivia Hart, Abhaya Dandekar, Anatoli Giritch, Somen Nandi, Yuri Gleba, Karen A. McDonald
Summary: The virus-based immunosorbent nanoparticle technology is a promising approach for biosensing and therapeutic antibody purification. It utilizes plant virions as chassis for diverse morphologies and high yield manufacturing. By consolidating the pro-vector system into a single vector, this study explores the potential of virus-based immunosorbent nanoparticles for therapeutic protein purification. The results demonstrate improved processing scheme and magnetic separation method for increased binding capacity and purity.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2022)
Article
Biotechnology & Applied Microbiology
Anne Chevrel, Leo Candela, Elisa Innocenti, Carolin Golibrzuch, Romas Skudas, Achim Schwaemmle, Manuel J. T. Carrondo, Olivier Kitten, Mikkel Nissum, Ricardo J. S. Silva
Summary: In this study, a novel custom affinity system based on Nanofitin protein scaffold was developed for the purification of biologics beyond antibodies. The system showed excellent performance in purifying a mutant form of streptolysin O, resulting in high purity product and improved process efficiency compared to the traditional three-step process. The custom affinity system has the potential to revolutionize the manufacturing of vaccines and other biological targets.
BIOTECHNOLOGY AND BIOENGINEERING
(2022)
Article
Multidisciplinary Sciences
Kristof Bozovicar, Barbara Jenko Bizjan, Anze Meden, Jernej Kovac, Tomaz Bratkovic
Summary: The development of a selective short linear peptide ligand of the IgG Fc region, capable of capturing all four IgG subclasses, was reported. This ligand was discovered through in vitro directed evolution and displayed high affinity for IgG, comparable to commercially employed protein A-based resin, after characterization and validation.
SCIENTIFIC REPORTS
(2021)
Article
Biochemistry & Molecular Biology
Julia Neumair, Claudia D'Ercole, Matteo De March, Martin Elsner, Michael Seidel, Ario de Marco
Summary: In this study, a new affinity chromatography platform composed of a macroporous epoxy monolith matrix functionalized with anti-CD63 nanobodies was developed. The platform successfully captured and released integral EVs from urine samples with negligible unspecific binding for circulating proteins. Size discrimination of eluted EVs was achieved by different elution approaches. The use of inexpensive recombinant nanobodies and scalable monolith materials make this platform feasible and affordable for large-scale production.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2023)
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)
Article
Biochemical Research Methods
Hesham Saeed, Eman Elsawy, Manal Shalaby, Manal Abdel-Fattah, Asmaa Hemida, Ahmad Eldoksh, Farid Shokry Ataya, Hesham Nematalla, Mohamed Elkewedi, Nikolaos N. Labrou, Nefertiti El-Nikhely
Summary: In this study, the recombinant L-ASNase from Dickeya chrysanthemi (DcL-ASNase) was successfully cloned, expressed, and purified, allowing for analysis of its properties and applications through techniques such as kinetics analysis and immunogenicity evaluation.
PREPARATIVE BIOCHEMISTRY & BIOTECHNOLOGY
(2022)
Article
Biology
Fereniki Perperopoulou, Nirmal Poudel, Anastassios C. Papageorgiou, Farid S. Ataya, Nikolaos E. Labrou
Summary: This study reported the kinetic and structural characterization of GSTM1-1 from Camelus dromedarius, revealing its narrow substrate specificity and limited ability to bind xenobiotic compounds. The crystal structures of CdGS tau M1-1 in complex with the substrate or the reaction product provided insights into the induced-fit catalytic mechanism. Furthermore, the thermodynamic stability of CdGSTM1-1 was investigated, showing its significant stabilization by ligands. These findings advance our understanding of the detoxification mechanisms and stress adaptation of camelid GSTs.
Article
Biochemistry & Molecular Biology
Charoutioun S. Bodourian, Nirmal Poudel, Anastassios C. Papageorgiou, Mariana Antoniadi, Nikolaos D. Georgakis, Hiroshi Abe, Nikolaos E. Labrou
Summary: This study evaluated the ligandability of human GSTM1-1 isoenzyme and identified pesticides that inhibit this enzyme. The results showed limited ligandability and ligand-binding promiscuity for GSTM1-1, with pirimicarb identified as the strongest inhibitor. The crystal structure of ligand-free GSTM1-1 was determined, revealing important structural features that contribute to ligand binding and specificity. These findings provide a basis for the design of new drugs targeting GSTM1-1.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2022)
Article
Biochemistry & Molecular Biology
Elisavet Ioannou, Anastassios C. Papageorgiou, Nikolaos E. Labrou
Summary: This study created a library of mutant GSTF genes through DNA shuffling, and identified enzyme variants with improved characteristics through activity screening and characterization. The structural and functional analysis of these enzyme variants revealed their involvement in crop herbicide resistance mechanisms.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2022)
Article
Biochemistry & Molecular Biology
Mohammed Hamed Alqarni, Ahmed Ibrahim Foudah, Magdy Mohamed Muharram, Aftab Alam, Nikolaos E. Labrou
Summary: The study found that myricetin can serve as a potential GST inhibitor, effectively inhibiting the activity of human glutathione transferase A1-1 and could be further developed as a natural, safe, and effective anti-cancer chemotherapy drug targeting GST.
Article
Biochemistry & Molecular Biology
Georgios E. Premetis, Angeliki Stathi, Anastassios C. Papageorgiou, Nikolaos E. Labrou
Summary: This study identified a potential endolysin (AbLys1) with high selectivity towards Acinetobacter baumannii and characterized its enzymatic activity. The structure of AbLys1 revealed two helical domains and short sequences with predicted antimicrobial activity. Engineering studies demonstrated the importance of these sequences in the lytic activity of AbLys1 towards A. baumannii cells.
Article
Biochemistry & Molecular Biology
Panagiota Pantiora, Veronika Furlan, Dimitris Matiadis, Barbara Mavroidi, Fereniki Perperopoulou, Anastassios C. Papageorgiou, Marina Sagnou, Urban Bren, Maria Pelecanou, Nikolaos E. Labrou
Summary: This study investigated the inhibitory effects of two curcuminoids and eleven monocarbonyl curcumin analogues against human glutathione transferase P1-1 (hGSTP1-1). The results showed that Demethoxycurcumin (Curcumin II) and three monocarbonyl curcumin analogues exhibited the highest inhibitory activity, with IC50 values ranging between 5.45 and 37.72 μM. The most potent inhibitors functioned as non-competitive/mixed-type inhibitors. Structural analysis revealed that the strongest hGSTP1-1 inhibitor, DM96, formed hydrophobic interactions and hydrogen bonds with specific residues, contributing to its superior inhibitory activity. The study highlights the potential of DM96 as a strong hGSTP1-1 inhibitor and its relevance for cancer chemosensitization.
Article
Biochemistry & Molecular Biology
Elisavet Ioannou, Nikolaos E. Labrou
Summary: Microalgae are a valuable source of enzymes and active ingredients. The proteolytic activity of Arthrospira platensis lysate was assessed and characterized, and a rapid and easy purification protocol was developed. The proteolytic activity of A. platensis lysate was used to develop a hydrogel formulation, which showed improved operational stability.
Article
Biochemistry & Molecular Biology
Evangelia G. Chronopoulou, Lana Mutabdzija, Nirmal Poudel, Anastassios C. Papageorgiou, Nikolaos E. Labrou
Summary: Glutathione transferases (GSTs) are versatile enzymes that primarily function in the detoxification of electrophilic compounds. A motif-directed redesign of human glutathione transferase A1-1 (hGSTA1-1) was performed, resulting in enzyme variants with enhanced catalytic activity and improved thermal stability. X-ray crystallographic analysis revealed the molecular basis for the effects of the mutations on enzyme stability and catalysis. This study provides valuable insights into the structure and function of alpha class GSTs.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2023)
Article
Biochemistry & Molecular Biology
Christina Varotsou, Georgios E. Premetis, Nikolaos E. Labrou
Summary: The rise of multidrug-resistant bacteria poses a significant threat to global public health. Exploiting phage endolysins shows promise as a solution to this problem. In this study, a N-acetylmuramoyl-L-alanine type-2 amidase from Propionibacterium bacteriophage PAC1 was characterized. The enzyme (PaAmi1) exhibited lytic activity against a range of human pathogens and was confirmed to degrade peptidoglycan. Engineered variants of PaAmi1 with antimicrobial peptides showed improved lytic activity against specific bacteria. These findings suggest that PaAmi1 is a potential antimicrobial agent and highlight the potential of phage genomes for designing novel endolysins.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2023)
Article
Biochemistry & Molecular Biology
Oleksii Kupreienko, Fotini Pouliou, Konstantinos Konstandinidis, Irene Axarli, Eleni Douni, Anastassios C. Papageorgiou, Nikolaos E. Labrou
Summary: Multidrug resistance poses a significant barrier to effective anticancer therapies. This study identified iprodione as a potential lead compound with high inhibitory potency against the isoenzyme GSTP1-1 from Mus musculus. The findings shed light on the inhibition mechanism of MmGSTP1-1 and provide a new compound for future drug development.
Article
Biochemistry & Molecular Biology
Nikolaos D. Georgakis, Elisavet Ioannou, Marianna Chatzikonstantinou, Marian Merino, Evangelia G. Chronopoulou, Jose Luis Mullor, Panagiotis Madesis, Nikolaos E. Labrou
Summary: In this study, the cosmeceutical properties of aqueous extracts from the microalgae strain Trachydiscus minutus were evaluated and compared with three widely used Chlorella strains. The T. minutus extracts showed the highest total antioxidant capacity and inhibitory potency towards elastase, indicating potential in controlling skin aging. The extracts also exhibited anti-inflammatory activity and protection against UVA-induced cell damage. The results suggest that T. minutus extract has promising cosmeceutical properties and potential for skincare use.
Review
Biochemistry & Molecular Biology
Alexandros Mavrommatis, Eleni Tsiplakou, Anastasia Zerva, Panagiota D. Pantiora, Nikolaos D. Georgakis, Georgia P. Tsintzou, Panagiotis Madesis, Nikolaos E. Labrou
Summary: Microalgae are a renewable source with beneficial effects on nutrition and health. They have been explored as feed additives in livestock animals, improving their health, product quality, and environmental impact. However, the cost-effective production and use of microalgae remains a challenge.
Article
Biotechnology & Applied Microbiology
Mohammed H. Alqarni, Ahmed Foudah, Aftab Alam, Mohammad A. Salkini, Magdy M. Muharram, Nikolaos E. Labrou, Pinki Rawat
Summary: Bacterial infections caused by antibiotic-resistant pathogens are a significant public health problem. Develop innovative strategies to effectively eliminate antibiotic-resistant microorganisms are urgently needed. Coumarin has shown significant effectiveness against MRSA and can be used as an alternative treatment. Nanoparticle-based targeted therapies can effectively treat MRSA infections.
BIOENGINEERING-BASEL
(2022)
Article
Polymer Science
Mohammed H. Alqarni, Ahmed Foudah, Aftab Alam, Mohammad A. Salkini, Magdy M. Muharram, Nikolaos E. Labrou, Piyush Kumar
Summary: This study aims to enhance the anti-fungal activity of rutin by encapsulating it in a nanocarrier system. The developed rutin-SNP gel showed significant activity against C. albicans skin candidiasis and was found to be biocompatible.
