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Phage display technology is a powerful tool for selecting monoclonal antibodies against a diverse set of antigens. Within toxinology, however, it remains challenging to generate monoclonal antibodies against many animal toxins, as they are difficult to obtain from venom. Recombinant toxins have been proposed as a solution to overcome this challenge, but so far, few have been used as antigens to generate neutralizing antibodies. Here, we describe the recombinant expression of α-cobratoxin in E. coli and its successful application as an antigen in a phage display selection campaign. From this campaign, an scFv (single-chain variable fragment) was isolated with similar binding affinity to a control scFv generated against the native toxin. The selected scFv recognizes a structural epitope, enabling it to inhibit the interaction between the acetylcholine receptor and the native toxin in vitro. This approach represents the first entirely in vitro antibody selection strategy for generating neutralizing monoclonal antibodies against a snake toxin.
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http://dx.doi.org/10.1016/j.nbt.2023.04.002 | 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 PDFBiotechnol Lett
September 2025
Department of Chemical Engineering, Hongik University, Sangsu-dong, Mapo-gu, Seoul, 04066, Republic of Korea.
The cell surface display system employs carrier proteins to present target proteins on the outer membrane of cells. This system enables functional proteins to be exposed on the exterior of living cells without cell lysis, allowing direct interaction with the surrounding environment. A major limitation of conventional approaches is the difficulty in displaying large-sized enzymes or antibodies, despite their critical roles in applications requiring functional domains that must remain intact, such as catalytic or antigen-binding sites.
View Article and Find Full Text PDFJ Control Release
September 2025
Laboratory of Precision and Nanomedicine, Institute of Biomedicine and Translational Medicine, University of Tartu, Ravila 14b, 50411 Tartu, Estonia; Materials Research Laboratory, University of California, Santa Barbara, CA 93106, USA. Electronic address:
Most chemotherapeutics distribute non-specifically throughout the body, resulting in off-target toxicities. Nanoparticle (NP) formulations provide a strategy to improve drug delivery by extending circulation time, protecting therapeutic agents from degradation, and enabling controlled release. However, delivering NPs effectively to solid tumors remains challenging due to the barriers within the tumor microenvironment.
View Article and Find Full Text PDFInt Immunopharmacol
September 2025
Key Laboratory for Biorheological Science and Technology of Chinese Ministry of Education, National Local Joint Engineering Lab for Vascular Implants, College of Bioengineering, Chongqing University, Chongqing 400044, China; JinFeng Laboratory, Chongqing, 401329, China. Electronic address: wanggx@cq
Thymic stromal lymphopoietin (TSLP) is an epithelial cell-derived cytokine that plays a crucial role in the pathophysiology of asthma, initiating multiple allergic cascade responses. Tezepelumab is the only monoclonal antibody currently approved for marketing, which acts by blocking TSLP binding to TSLPR. However, it is reported that a TSLP trap which simultaneously block TSLP binding with TSLPR and IL-7Rα has better efficiency in the repression of TSLP signal pathway.
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.
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