Article
Biotechnology & Applied Microbiology
Nafiseh Sanei Ata-Abadi, Mahboobeh Forouzanfar, Kianoush Dormiani, Shiva Rouhollahi Varnosfaderani, Leila Pirjamali, Mohammad Hossein Nasr-Esfahani, Reza Moradi Hajidavaloo
Summary: In this study, we successfully constructed stable cell lines in HEK293T cells using the PhiC31 integrase system, which can efficiently produce and secrete active rhPH20. Quantitative and functional analysis showed that the secreted rhPH20 had specific activity and could remove cumulus cells from oocytes. Additionally, the results demonstrated the ability of the PhiC31 enzyme to insert the PH20-expressing donors into specific pseudo attP sites in the genome of target cells with different copy numbers.
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
(2022)
Article
Biotechnology & Applied Microbiology
Sana Pourtabatabaei, Samaneh Ghanbari, Narges Damavandi, Elham Bayat, Mozhgan Raigani, Sirous Zeinali, Fatemeh Davami
Summary: Chinese hamster ovary (CHO) cells are widely used for manufacturing therapeutic proteins, and in this study, stable cell lines were successfully generated with targeted integration of reporter genes using the CRISPR/Cas9 platform. The study showed that higher efficiency and expression homogeneity were observed in the pseudo attP site associated with chromosome 6 compared to the pseudo attP site from chromosome 3.
JOURNAL OF BIOTECHNOLOGY
(2021)
Article
Biochemical Research Methods
Weidu Song, Chuanhao Zhang, Huichao Lin, Taiyi Zhang, Haixia Liu, Xiaowen Huang
Summary: The proposed real-time rotary PCR reaction system (RRP) meets the requirements of low cost, low-power consumption, potential for miniaturization, and portability. Experimental and theoretical results demonstrate its feasibility in the rapid quantitative detection of Pseudomonas aeruginosa and have broad application prospects in bacteria detection and food safety.
Article
Biochemical Research Methods
Shaolei Huang, Yiquan An, Bangchao Xi, Xianglian Gong, Zhongfu Chen, Shan Shao, Shengxiang Ge, Jun Zhang, Dongxu Zhang, Ningshao Xia
Summary: Nucleic acid detection is a method that directly identifies the presence of pathogenic microorganisms and has advantages of high sensitivity, commendable specificity, and a short window period. A real-time PCR system based on multiple temperature zones was proposed to solve the time-consuming problem, enabling fast detection speed and high sensitivity under extreme conditions.
Article
Multidisciplinary Sciences
Jae Jong Kim, Hyoung-Min Park, A. Young Kyoung, In Kyung Park, Si-Kyu Lim, Byoung Chul Park
Summary: Genetic mutations like SNP are common in genetic disorders and cancers, and PCR methods are widely used for SNP detection. The specifically designed PCR platform STexS can efficiently distinguish small amounts of SNP mixed in normal DNA, aiding in targeting potential SNPs or genetically mutated biomarkers in human clinical samples.
SCIENTIFIC REPORTS
(2021)
Article
Chemistry, Analytical
Lin Sun, Jingli Yan, Yilin Liu, Ruipeng Wang, Yongqiang Cheng
Summary: 5-hydroxymethylcytosine (5hmC) is closely correlated with various diseases. This study proposes a chemical oxidation-free and site-specific 5hmC assay, which has high selectivity and sensitivity. The method can simultaneously measure the content and percentage of 5hmC.
SENSORS AND ACTUATORS B-CHEMICAL
(2022)
Article
Medicine, Research & Experimental
Xiaozhen Hong, Yanling Ying, Jingjing Zhang, Shu Chen, Xianguo Xu, Ji He, Faming Zhu
Summary: Three novel splice site variations in the ABO gene were identified in Chinese individuals, resulting in decreased A or B antigen expression and the formation of ABO subtypes.
JOURNAL OF TRANSLATIONAL MEDICINE
(2021)
Article
Chemistry, Multidisciplinary
Sudheendran Mavila, Heidi R. Culver, Alex J. Anderson, Tania R. Prieto, Christopher N. Bowman
Summary: An athermal approach using self-immolative thioester linked nucleic acids (TENA) for mRNA enrichment from total RNA has shown comparable performance to a commercial kit. The method is compatible with downstream applications such as RT-qPCR for gene expression analysis.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2022)
Article
Ophthalmology
Satoko Nakano, Yasuhiro Tomaru, Toshiaki Kubota, Hiroshi Takase, Manabu Mochizuki, Norio Shimizu, Sunao Sugita
Summary: The Direct Strip PCR test offers rapid and accurate detection of infectious pathogens causing uveitis. It shows high concordance with qPCR results, even when used by beginners, indicating its reliability for clinical use. Additionally, it can be used for intraoperative diagnosis, improving the prognosis of infectious diseases.
Article
Medicine, General & Internal
Jeong-Hwa Kim, Jae-Woon Oh, Young Lee, Jeong-Ho Yun, Seong-Ho Choi, Dong-Woon Lee
Summary: This study found that detecting 18 bacterial species in mouth-rinse samples can be used to diagnose periodontitis, with Treponema denticola, Prevotella nigrescens, and Streptococcus mitis significantly associated with severe periodontitis. Further research is needed to assess the potential of oral-rinsing solutions in indicating oral infection risk and improving oral hygiene.
JOURNAL OF CLINICAL MEDICINE
(2021)
Article
Parasitology
Gereon Schares, Majda Globokar Vrhovec, Mareen Tuschy, Maike Joeres, Andrea Baerwald, Bretislav Koudela, Jitender P. Dubey, Pavlo Maksimov, Franz J. Conraths
Summary: The 529-bp repeat of H. hammondi can be used to establish a quantitative real-time PCR assay, with excellent specificity and sensitivity. This assay is a valuable diagnostic tool for epidemiological and cell biological studies on H. hammondi and related parasites.
PARASITES & VECTORS
(2021)
Article
Multidisciplinary Sciences
Ronaldo Celerino da Silva, Suelen Cristina de Lima, Wendell Paloma Maria dos Santos Reis, Jurandy Junior Ferraz de Magalhaes, Ronaldo Nascimento de Oliveira Magalhaes, Brijesh Rathi, Alain Kohl, Marcos Andre Cavalcanti Bezerra, Lindomar Pena
Summary: This study compared three DNA extraction methods for COVID-19 host genetic studies and found that Chelex (R) 100 resin is a cheap, simple, fast, and high-quality method for extracting DNA from nasopharyngeal samples of COVID-19 patients.
Article
Veterinary Sciences
Gilad Segev, Tal Yaaran, Sarah Maurice, Gad Baneth
Summary: qRT-PCR is sensitive for detecting CPV-2 infection, but its specificity is temporarily hampered during the vaccination period. Blood, feces, and pharynx are suitable sampling sites. Fecal samples have the lowest sensitivity in sick dogs, while showing the highest positivity in puppies after vaccination.
JOURNAL OF VETERINARY INTERNAL MEDICINE
(2022)
Article
Biotechnology & Applied Microbiology
Honggi Min, Seul Mi Kim, Dongwoo Kim, Solhwi Lee, Sumin Lee, Jae Seong Lee
Summary: In this study, a hybrid cell line development platform was developed to increase the productivity of targeted integrants through the amplification of transgene copies. The results showed that targeted integrants exhibited a 3.6-fold increase in EGFP expression and increased copy numbers of DHFR and EGFP when exposed to 200 nM MTX. A single-step MTX amplification increased the specific monoclonal antibody productivity by 2.8-fold. The study provides a new strategy for increasing the productivity of CHO cell lines.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2022)
Article
Optics
Damian Presti, Julian Bergier, Lucas Ripoll, Cristina Borio, Gustavo A. Torchia, Marcos Bilen
Summary: Since the global pandemic of SARS-CoV-2, the need for a low-cost and faster diagnostic tool for evaluating symptomatic patients has emerged. Point-of-care testing and point-of-need testing systems have been developed to provide accurate and rapid diagnostics. In this study, a bio-photonic device using an iso-thermal system was developed for detecting SARS-CoV-2. The device showed comparable sensitivity to the commercially used reference method and was built with simple and low-cost components, making it a high-efficiency and affordable instrument.
JOURNAL OF THE OPTICAL SOCIETY OF AMERICA A-OPTICS IMAGE SCIENCE AND VISION
(2023)
Article
Multidisciplinary Sciences
Zohreh Sharifi, Faezeh Rahiminejad, Atefeh Joudaki, Ameneh Sarhadi Bandehi, Hossein Farahzadi, Yeganeh Keshvar, Fatemeh Golnabi, Sanaz Naderi, Rasaneh Yazdani, Mehdi Shafaat, Shirin Ghadami, Maryam Abiri, Sirous Zeinali
SCIENTIFIC REPORTS
(2019)
Article
Obstetrics & Gynecology
Yeganeh Keshvar, Solmaz Sabeghi, Zohreh Sharifi, Kiyana Sadat Fatemi, Panti Fouladi, Shahrzad Younesi Khah, Faezeh Rahiminejad, Atefeh Joudaki, Masoume Amini, Hamideh Bagherian, Marefat Ghaffari Novin, Mansoureh Movahedin, Marzieh Mojbafan, Sirous Zeinali
Summary: The study reports the use of preimplantation genetic diagnosis (PGD) for diagnosing beta thalassemia and evaluating chromosomal aneuploidies, HLA typing, and sex selection. The results show successful detection using STR markers and a high success rate in pregnancies and live births. PGD is a powerful diagnostic tool for carrier couples who desire a healthy child and wish to avoid medical abortion.
BMC PREGNANCY AND CHILDBIRTH
(2022)
Article
Biochemistry & Molecular Biology
Samira Bushra, Ying-Ni Lin, Atefeh Joudaki, Mikako Ito, Bisei Ohkawara, Kinji Ohno, Akio Masuda
Summary: Agrin is a crucial heparan sulfate proteoglycan that clusters acetylcholine receptors at the neuromuscular junction. The inclusion of Y and Z exons in neuronal isoforms of agrin is regulated by binding sites for PTBP1, which were found to be enriched in the human AGRN gene. PTBP1-silencing enhanced the coordinated inclusion of Y and Z exons, while deletion analysis identified five PTBP1-binding sites with significant splicing repression activities. Neuronal differentiation downregulates PTBP1 expression and promotes the coordinated inclusion of Y and Z exons, suggesting that the reduction of the PTBP1-RNA network is essential for generating neuron-specific agrin isoforms.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2023)
Article
Andrology
S. Bahrami Zadegan, S. Dabbagh Bagheri, A. Joudaki, M. H. Samiee Aref, A. H. Saeidian, M. Abiri, S. Zeinali
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)
