Article
Biochemical Research Methods
Andrei Hutanu, David Boelsterli, Claudio Schmidli, Cristina Montealegre, Mike H. N. Dang Thai, Balazs Bobaly, Marius Koch, Maria A. Schwarz
Summary: With FDA approval of rAAV-based gene therapies, efforts are being made to improve manufacturing and quality control processes. Various analytical methods are used to analyze the complex DNA composition inside rAAVs, showing that a combination of techniques is essential for a clear understanding of the virus interior.
Letter
Biochemical Research Methods
Martin Bartas, Pratik Goswami, Matej Lexa, Jiri Cerven, Adriana Volna, Miroslav Fojta, Vaclav Brazda, Petr Pecinka
Summary: The study found that mutations in the new SARS-CoV-2 strains are significantly enriched in inverted repeat loci.
BRIEFINGS IN BIOINFORMATICS
(2021)
Article
Biochemistry & Molecular Biology
Ante Turudic, Zlatko Liber, Martina Grdisa, Jernej Jakse, Filip Varga, Zlatko Satovic
Summary: The development of next-generation sequencing technology and the increasing amount of sequencing data have made bioinformatic tools used in genome assembly important. In the final step of genome annotation, specialized tools are used to identify organelle genes and structural features, with most tools targeting chloroplast sequences.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2022)
Article
Biotechnology & Applied Microbiology
Hiroaki Oyama, Kentaro Ishii, Takahiro Maruno, Tetsuo Torisu, Susumu Uchiyama
Summary: Recombinant adeno-associated virus (AAV) is an important tool in human gene therapy, and this study identified two VP3-related components in AAV vectors using sodium dodecyl sulfate-polyacrylamide gel electrophoresis, capillary gel electrophoresis, and liquid chromatography-UV-mass spectrometry. The study suggests that complementary evaluation of VP components using CGE and LC-UV-MS methods is necessary for accurate quantification and understanding of VP stoichiometry in AAV vectors.
HUMAN GENE THERAPY
(2021)
Article
Microbiology
Perumal Arumugam Desingu, K. Nagarajan, Nagalingam R. Sundaresan
Summary: Different groups of MPXVs contain unique tandem repeats in the ITR regions, which may play a role in the genetic diversity of the virus. These repeats are only found in MPXVs and are different from the tandem repeats in human and rodent genomes.
MICROBIOLOGY SPECTRUM
(2023)
Article
Microbiology
Yucheng Xia, Huanyu Cheng, Jiang Zhong
Summary: Mimivirus is a type of DNA virus infecting amoebas, with a genome encoding 1,062 predicted ORFs and two identical inverted terminal repeats. A new member of Mimivirus lineage C was isolated from a fresh water pond in Shanghai, China, and its complete genome was reported in this study. The discovery of two ITRs in the Mimivirus genome provides new insights into its structure.
FRONTIERS IN MICROBIOLOGY
(2022)
Article
Biochemistry & Molecular Biology
Xiaomin Zhang, Ge Yang, Wenjing Liu, Qing Liu, Zhuoran Wang, Kelong Fan, Feng Qu, Yuanyu Huang
Summary: Using CE-SELEX, four candidate aptamers with a stem-loop structure were selected targeting LRP6 protein. Among them, AptLRP6-A3 showed the highest affinity with LRP6-overexpressed human breast cancer cells. This LRP6 aptamer holds promise for rapid diagnosis and treatment of LRP6-related diseases.
Article
Multidisciplinary Sciences
Andrew Prantner, Dianna Maar
Summary: In this study, a simple and robust ddPCR protocol was developed for AAV characterization, which involved dilution of AAV into the ddPCR concentration range, thermal capsid lysis, and amplification of various regions of the AAV vector genome. Furthermore, different methods of genome integrity analysis were compared, and it was found that determining integrity based on the excess number of positive droplets yielded expected values, while using the double positive droplet percentage produced inconsistent results.
Article
Chemistry, Multidisciplinary
Samuele Zoratto, Victor U. Weiss, Gernot Friedbacher, Carsten Buengener, Robert Pletzenauer, Alexandra Foettinger-Vacha, Michael Graninger, Guenter Allmaier
Summary: The study presents a method to analyze purified AAV8 batches using nES GEMMA technology, along with AF4 and AFM techniques as orthogonal methods for VLP characterization. By combining these techniques, a comprehensive characterization of AAV8 VLPs can be achieved.
Article
Chemistry, Analytical
Robert Farsang, Noemi Kovacs, Marton Szigeti, Hajnalka Jankovics, Ferenc Vonderviszt, Andras Guttman
Summary: This paper reports the design and production of 6HIS-tagged exoglycosidases for efficient solid phase N-glycan sequencing. The enzymes were generated in bacterial expression systems and immobilized via the 6HIS-tag for high digestion performance. The immobilized enzymes were demonstrated to have high efficiency in digesting glycoproteins through analysis of commercial glycoprotein therapeutic and fluorophore labeled glycans in human serum.
ANALYTICA CHIMICA ACTA
(2022)
Article
Chemistry, Analytical
Bo Liu, Wentao Wang, Tie Gao, Lu Huang, Huihong Fan, Hong-Xu Chen
Summary: Cobratide, a peptide drug extracted from Chinese cobra venom, has shown potential in treating not only chronic pain but also COVID-19. A protocol combining sheathless CE-MS and CE-UV methods was established for the separation, identification, and quantification of cobratide and its impurities, providing valuable information for quality control and process analysis of peptide drugs.
ANALYTICAL METHODS
(2021)
Article
Medicine, General & Internal
Katarzyna Pauter, Viorica Railean-Plugaru, Michal Zloch, Pawel Pomastowski, Malgorzata Szultka-Mlynska, Boguslaw Buszewski
Summary: The study aimed to characterize biofilm before and after antibiotic treatment using methods such as 16S rRNA gene sequencing and capillary electrophoresis. Results showed that antibiotics (amoxicillin and gentamicin) can alter the surface charge of bacterial cells, leading to cell clumping. Additionally, microscopic and spectrometry studies revealed that antibiotics inhibit cell wall synthesis and ribosomal protein synthesis in bacterial cells.
JOURNAL OF CLINICAL MEDICINE
(2022)
Review
Chemistry, Analytical
Boguslaw Buszewski, Ewelina Maslak, Michal Zloch, Viorica Railean-Plugaru, Ewa Klodzinska, Pawel Pomastowski
Summary: Fast determination and identification of pathogens is crucial in various fields, especially during pandemics like COVID-19. Innovative methods combining electromigration techniques and laser desorption/ionization analysis can simultaneously analyze the proteome, lipidome, and genome of pathogens.
TRAC-TRENDS IN ANALYTICAL CHEMISTRY
(2021)
Article
Food Science & Technology
Yu-Min Lee, Shinyoung Lee, Hae-Yeong Kim
Summary: With increased consumption of seafood products, there have been continuous reports of food fraud involving fish resources, particularly codfish. A multiplex PCR method was developed in this study to simultaneously identify five cod species within Gadidae, successfully applied to 40 commercial food products. This detection method will efficiently manage seafood authentication by analyzing multiple cod species simultaneously.
Article
Chemistry, Analytical
Pingjing Li, Lulu Wang, Rui Guo, Huatao Feng, Ya Ji, Si Ying Lim, Bao Hui Ng, Anna Karen Carrasco Laserna, Shaheer Khan, Shiaw-Min Chen, Sam Fong Yau Li
Summary: Recombinant protein biopharmaceuticals are important in current drug development, and the glycosylation profile of these proteins plays a crucial role in their quality. This study developed a CE-LIF method for accurate glycan identification by cross-identification using two capillary coatings and three labeling dyes. The use of homemade PVA and commercial coatings showed similar separation profiles, and the combination of fluorescent dyes provided better accuracy.