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Phage display of combinatorial antibody libraries is a versatile tool in the field of antibody engineering, with diverse applications including monoclonal antibody (mAb) discovery, affinity maturation, and humanization. To improve the selection efficiency of antibody libraries, we developed a new phagemid display system that addresses the complication of bald phage propagation. The phagemid facilitates the biotinylation of fragment of antigen binding (Fab) antibody fragments displayed on phage via Sortase A catalysis and the subsequent enrichment of Fab-displaying phage during selections. In multiple contexts, this selection approach improved the enrichment of target-reactive mAbs by depleting background phage. Panels of cancer cell line-reactive mAbs with high diversity and specificity were isolated from a naïve chimeric rabbit/human Fab library using this approach, highlighting its potential to accelerate antibody engineering efforts and to empower concerted antibody drug and target discovery.
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http://dx.doi.org/10.1016/j.jmb.2018.09.003 | DOI Listing |
J Biol Chem
September 2025
Key Laboratory of Systems Health Science of Zhejiang Province, School of Life Science, Hangzhou Institute for Advanced Study, Hangzhou 310024; University of Chinese Academy of Sciences, China. Electronic address:
Phage display libraries of human single-chain variable fragments (scFv) serve as a valuable resource for generating fully human antibodies for scientific and clinical applications. In this study, we designed and constructed a highly diverse semi-synthetic humanized scFv phage display library using an optimized Kunkel mutagenesis approach. Our optimizations eliminated residual template, enhancing mutagenesis efficiency and expanding library diversity with a reservoir capacity exceeding 10.
View Article and Find Full Text PDFArch Microbiol
September 2025
Department of Infectious Disease, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, No. 639, Zhizaoju Road, Huangpu District, Shanghai, 200011, China.
Highly pathogenic avian influenza (HPAI) H5N1 virus poses a continuing global public health threat due to its outbreaks in poultry farms and zoonotic transmission from birds to humans. In the quest of effective therapeutics against H5N1 infection, antibodies with broad neutralizing activity have attracted significant attention. In this study, we employed a phage display technique to select and identify VHH antibodies with specific neutralizing activity against H5N1 hemagglutinin (HA) from an immune llama-derived antibody library.
View Article and Find Full Text PDFJ Neurol
September 2025
Department of Neurology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, 100730, China.
Objective: To evaluate the clinical efficacy of tocilizumab, a interleukin-6 (IL-6) receptor blocker, for the treatment of acute necrotizing encephalopathy (ANE).
Methods: PubMed, Cochrane Library, Embase, and Web of Science were searched for systematic review based on PRISMA guidelines. ANE patients treated with and without tocilizumab were included.
Int Immunopharmacol
September 2025
Center for Genetic Engineering and Biotechnology, Cubanacan, 10600 Havana, POBox 6162, Cuba. Electronic address:
Vascular endothelial growth factor (VEGF) is a key player in the development and progression of several diseases, most notably cancer and retinal disorders. Over the last twenty years, VEGF has emerged as a significant therapeutic target for these conditions. This study reports the isolation and characterization of a fully synthetic, humanized, affinity-matured single-domain antibody fragment (VHH) designed to target VEGF.
View Article and Find Full Text PDFACS Synth Biol
September 2025
Jiangsu Key Laboratory for Food Quality and Safety, State Key Laboratory Cultivation Base of Ministry of Science and Technology, Institute of Food Safety and Nutrition, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China.
Anti-idiotypic antibodies (Anti-Ids) possess the properties to mimic the structure and biological activity of an antigen, which can be utilized for preventing and monitoring hazards. In this study, Nb4Mutant6-Anti-Id, which mimics the structure and antibacterial activity of vancomycin, was designed based on phage display antibody library screening and mutagenesis technology. The affinity of Nb4Mutant6-Anti-Id for the coated antigens of Van-pAbs F(ab)2 and inactivated cells was 6.
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