C-Terminal Cysteine PEGylation of Adalimumab Fab with an Engineered Interchain SS Bond
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- Nakamura Hitomi
- Faculty of Pharmaceutical Sciences, Sojo University
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- Anraku Makoto
- Faculty of Pharmaceutical Sciences, Sojo University
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- Oda-Ueda Naoko
- Faculty of Pharmaceutical Sciences, Sojo University
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- Ueda Tadashi
- Graduate School of Pharmaceutical Sciences, Kyushu University
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- Ohkuri Takatoshi
- Faculty of Pharmaceutical Sciences, Sojo University
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<p>Conjugation with polyethylene glycol (PEG) is performed to increase serum half-life of the Fab for clinical applications. However, current designs for recombinant Fab only allow PEGylation at the interchain SS bond (disulfide bond) at the C-terminal end of the heavy chain and light chain of the Fab, which the decrease of thermostability occurred by partial reduction of the interchain SS bond. An adalimumab Fab mutant with a novel interchain SS bond (CH1 : C177–CL : C160) and one cysteine at the C-terminal end (mutSS FabSH) was designed to maintain Fab thermostability and for site-specific PEGylation. MutSS FabSH was expressed in Pichia pastoris and purified mutSS FabSH was conjugated with 20-kDa PEG targeted at the free cysteine. Based on enzyme-linked immunosorbent assay (ELISA), PEGylation did not affect the binding capacity of the mutSS FabSH. To confirm the influence of PEGylation on the pharmacokinetic behavior of the Fab, PEGylated mutSS FabSH was administered to rats via tail vein injection. Analysis of the mean serum concentration of the PEGylated mutSS FabSH versus time through ELISA indicated an increase in half-life compared to that of non-PEGylated wild-type Fab. Consequently, we have successfully demonstrated that a Fab mutant with a novel interchain SS bond and one free cysteine at the C-terminal end can be PEGylated without changes in functionality. This design can potentially be used as a platform for modification of other recombinant Fabs.</p>
収録刊行物
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- Biological & Pharmaceutical Bulletin
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Biological & Pharmaceutical Bulletin 43 (3), 418-423, 2020-03-01
公益社団法人 日本薬学会
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詳細情報 詳細情報について
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- CRID
- 1390565134832142464
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- NII論文ID
- 130007804448
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- NII書誌ID
- AA10885497
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- ISSN
- 13475215
- 09186158
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- NDL書誌ID
- 030274804
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- PubMed
- 31866612
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- 本文言語コード
- en
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- データソース種別
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- JaLC
- NDL
- Crossref
- PubMed
- CiNii Articles
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