Convenient method of producing cyclic single-chain Fv antibodies by split-intein-mediated protein ligation and chaperone co-expression
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<jats:title>Abstract</jats:title> <jats:p>Single-chain Fv (scFv) is a recombinant antibody in which the variable regions of the heavy chain (VH) and light chain (VL) are connected by a short flexible polypeptide linker. Compared with monoclonal antibodies, scFvs have the advantages of low-cost production using Escherichia coli and easy genetic manipulation. ScFvs are, therefore, regarded as useful modules for producing next-generation medical antibodies. The practical use of scFvs has been limited due to their aggregation propensity mediated by interchain VH–VL interactions. To overcome this problem, we recently reported a cyclic scFv whose N-terminus and C-terminus were connected by sortase A-mediated ligation. Preparation of cyclic scFv is, however, a time-consuming process. To accelerate the application study of cyclic scFv, we developed a method to produce cyclic scFv by the combined use of a protein ligation technique based on protein trans-splicing reaction (PTS) by split intein and a chaperone co-expression system. This method allows for the preparation of active cyclic scFv from the cytoplasm of E. coli. The present method was applied to the production of cyclic 73MuL9-scFv, a GA-pyridine antibody, as a kind of advanced glycation end-product. These findings are expected to evoke further application study of cyclic scFv.</jats:p>
収録刊行物
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- The journal of biochemistry
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The journal of biochemistry 168 (3), 257-263, 2020-09
Tokyo : Japanese Biochemical Society
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詳細情報 詳細情報について
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- CRID
- 1523106604812320128
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- NII論文ID
- 40022375418
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- NII書誌ID
- AA00694073
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- ISSN
- 0021924X
- 17562651
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- NDL書誌ID
- 030687187
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- 本文言語コード
- en
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- NDL 雑誌分類
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- ZR2(科学技術--生物学--生化学)
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- データソース種別
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- NDL
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