Next-Generation Sequencing and Proteomics-Enabled Approach for Rapid and High-Throughput Isolation of Virus-Neutralizing Nanobodies.
| Autor: | Wu X; State Key Laboratory of Pharmaceutical Biotechnology, Medical School, Nanjing University, Nanjing, China.; Chemistry and Biomedicine Innovation Center, Jiangsu Key Laboratory of Molecular Medicine, Medical School, Nanjing University, Nanjing, China., Zhang J; State Key Laboratory of Pharmaceutical Biotechnology, Medical School, Nanjing University, Nanjing, China.; Chemistry and Biomedicine Innovation Center, Jiangsu Key Laboratory of Molecular Medicine, Medical School, Nanjing University, Nanjing, China., Fang L; State Key Laboratory of Pharmaceutical Biotechnology, Medical School, Nanjing University, Nanjing, China. njfanglei@nju.edu.cn.; Chemistry and Biomedicine Innovation Center, Jiangsu Key Laboratory of Molecular Medicine, Medical School, Nanjing University, Nanjing, China. njfanglei@nju.edu.cn. |
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| Jazyk: | angličtina |
| Zdroj: | Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2024; Vol. 2824, pp. 135-146. |
| DOI: | 10.1007/978-1-0716-3926-9_10 |
| Abstrakt: | Compared with traditional antibodies, nanobodies from camelids have various advantages, including small molecular weight, high affinity, low immunogenicity, convenient production through genetic engineering, etc. Here we combined next-generation sequencing (NGS) with proteomics technology based on affinity purification-mass spectrometry (AP-MS) and bioinformatics analysis to high-throughput screen monoclonal nanobodies from camels immunized with surface glycoprotein (glycoprotein N, Gn) of severe fever with thrombocytopenia syndrome virus and fulfilled production of the screened anti-Gn monoclonal nanobody with high affinity by genetic engineering. The innovative high-throughput technical route developed here could also be expanded to the production of neutralizing nanobodies specific for Rift Valley fever virus. (© 2024. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.) |
| Databáze: | MEDLINE |
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