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Isolation of PCSK9-specific nanobodies from synthetic libraries using a combined protein selection strategy.
Sci Rep
January 2025
Biological Engineering Program, Faculty of Engineering, King Mongkut's University of Technology Thonburi, Bangkok, 10140, Thailand.
Nanobodies (Nbs) hold great potential to replace conventional antibodies in various biomedical applications. However, conventional methods for their discovery can be time-consuming and expensive. We have developed a reliable protein selection strategy that combines magnetic activated cell sorting (MACS)-based screening of yeast surface display (YSD) libraries and functional ligand-binding identification by Tat-based recognition of associating proteins (FLI-TRAP) to isolate antigen-specific Nbs from synthetic libraries.
View Article and Find Full Text PDFCloning a Chloroplast Genome in and .
Bio Protoc
January 2025
Biochemistry Department, Western University, London, Canada.
Chloroplast genomes present an alternative strategy for large-scale engineering of photosynthetic eukaryotes. Prior to our work, the chloroplast genomes of (204 kb) and (140 kb) had been cloned using bacterial and yeast artificial chromosome (BAC/YAC) libraries, respectively. These methods lack design flexibility as they are reliant upon the random capture of genomic fragments during BAC/YAC library creation; additionally, both demonstrated a low efficiency (≤ 10%) for correct assembly of the genome in yeast.
View Article and Find Full Text PDFCellREADR: An ADAR-based RNA sensor-actuator device.
Methods Enzymol
January 2025
Department of Neurobiology, Duke University School of Medicine, Durham, NC, United States; Department of Biomedical Engineering, Duke University, Durham, NC, United States. Electronic address:
RNAs are central mediators of genetic information flow and gene regulation that underlie diverse cell types and cell states across species. Thus, methods that can sense and respond to RNA profiles in living cells will have broad applications in biology and medicine. CellREADR - Cell access through RNA sensing by Endogenous ADAR (adenosine deaminase acting on RNA), is a programmable RNA sensor-actuator technology that couples the detection of a cell-defining RNA to the translation of an effector protein to monitor and manipulate the cell.
View Article and Find Full Text PDFRecombinant Abelson Tyrosine Protein Kinase (rAbl) Regulates Various Functions of Buffalo Peripheral Blood Mononuclear Cells.
Animals (Basel)
January 2025
Guangxi Colleges and Universities Key Laboratory of Prevention and Control for Animal Disease, College of Animal Science and Technology, Guangxi University, Nanning 530005, China.
can modulate host immune mechanisms through excretory-secretory products (ESP). As one of the components of ESP, it is unknown whether Abelson tyrosine protein kinase (Abl) is involved in parasite-host immune interaction. To investigate the immunoregulatory function of Abl in , we cloned and expressed the gigantica Abl protein and assessed its effect on specific immune functions of buffalo peripheral blood mononuclear cells (PBMCs).
View Article and Find Full Text PDFMonoclonal antibody development and antigenic epitope identification of chicken pro-IL-1β.
Poult Sci
January 2025
Avian Immunosuppressive Diseases Division, State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, the Chinese Academy of Agricultural Sciences, Harbin, 150069, China; Jiangsu Co-Innovation Center for the Prevention and Control of Important Animal Infectious Disease and Zoonosis, Yangzhou University, Yangzhou, 225009, China. Electronic address:
Pro-IL-1β is an important inflammatory factor and is also a biomarker for detecting early pro-inflammatory immune responses. However, commercially available antibodies against chicken inflammatory factors are lacking, which prohibits an in-depth exploration of the mechanism of chicken inflammation. This study cloned and expressed chicken pro-IL-1β, and developed a hybridoma cell line 1E12 capable of stably secreting chicken pro-IL-1β monoclonal antibody (mAb).
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