Production of antibodies and antibody fragments containing non-natural amino acids in Escherichia coli
Therapeutic bioconjugates are emerging as an essential tool to combat human disease. Site-specific conjugation technologies are widely recognized as the optimal approach for producing homogeneous drug products. Non-natural amino acid (nnAA) incorporation allows the introduction of bioconjugation han...
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2024-12-01
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| Series: | mAbs |
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| Online Access: | https://www.tandfonline.com/doi/10.1080/19420862.2024.2316872 |
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| author | Jacquelyn Blake-Hedges Dan Groff Wilson Foo Jeffrey Hanson Elenor Castillo Miao Wen Diana Cheung Mary Rose Masikat Jian Lu Young Park Nina Abi Carlos Hans Usman Kevin Fong Abigail Yu Sihong Zhou Joyce Kwong Cuong Tran Xiaofan Li Dawei Yuan Trevor Hallam Gang Yin |
| author_facet | Jacquelyn Blake-Hedges Dan Groff Wilson Foo Jeffrey Hanson Elenor Castillo Miao Wen Diana Cheung Mary Rose Masikat Jian Lu Young Park Nina Abi Carlos Hans Usman Kevin Fong Abigail Yu Sihong Zhou Joyce Kwong Cuong Tran Xiaofan Li Dawei Yuan Trevor Hallam Gang Yin |
| author_sort | Jacquelyn Blake-Hedges |
| collection | DOAJ |
| description | Therapeutic bioconjugates are emerging as an essential tool to combat human disease. Site-specific conjugation technologies are widely recognized as the optimal approach for producing homogeneous drug products. Non-natural amino acid (nnAA) incorporation allows the introduction of bioconjugation handles at genetically defined locations. Escherichia coli (E. coli) is a facile host for therapeutic nnAA protein synthesis because it can stably replicate plasmids encoding genes for product and nnAA incorporation. Here, we demonstrate that by engineering E. coli to incorporate high levels of nnAAs, it is feasible to produce nnAA-containing antibody fragments and full-length immunoglobulin Gs (IgGs) in the cytoplasm of E. coli. Using high-density fermentation, it was possible to produce both of these types of molecules with site-specifically incorporated nnAAs at titers > 1 g/L. We anticipate this strategy will help simplify the production and manufacture of promising antibody therapeutics. |
| format | Article |
| id | doaj-art-1e674937a64d456a8d216791a8a935ac |
| institution | Kabale University |
| issn | 1942-0862 1942-0870 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | mAbs |
| spelling | doaj-art-1e674937a64d456a8d216791a8a935ac2025-01-31T04:19:38ZengTaylor & Francis GroupmAbs1942-08621942-08702024-12-0116110.1080/19420862.2024.2316872Production of antibodies and antibody fragments containing non-natural amino acids in Escherichia coliJacquelyn Blake-Hedges0Dan Groff1Wilson Foo2Jeffrey Hanson3Elenor Castillo4Miao Wen5Diana Cheung6Mary Rose Masikat7Jian Lu8Young Park9Nina Abi Carlos10Hans Usman11Kevin Fong12Abigail Yu13Sihong Zhou14Joyce Kwong15Cuong Tran16Xiaofan Li17Dawei Yuan18Trevor Hallam19Gang Yin20Research and Process Development, Sutro Biopharma, Inc, South San Francisco, CA, USAResearch and Process Development, Sutro Biopharma, Inc, South San Francisco, CA, USAResearch and Process Development, Sutro Biopharma, Inc, South San Francisco, CA, USAResearch and Process Development, Sutro Biopharma, Inc, South San Francisco, CA, USAResearch and Process Development, Sutro Biopharma, Inc, South San Francisco, CA, USAResearch and Process Development, Sutro Biopharma, Inc, South San Francisco, CA, USAResearch and Process Development, Sutro Biopharma, Inc, South San Francisco, CA, USAResearch and Process Development, Sutro Biopharma, Inc, South San Francisco, CA, USAResearch and Process Development, Sutro Biopharma, Inc, South San Francisco, CA, USAResearch and Process Development, Sutro Biopharma, Inc, South San Francisco, CA, USAResearch and Process Development, Sutro Biopharma, Inc, South San Francisco, CA, USAResearch and Process Development, Sutro Biopharma, Inc, South San Francisco, CA, USAResearch and Process Development, Sutro Biopharma, Inc, South San Francisco, CA, USAResearch and Process Development, Sutro Biopharma, Inc, South San Francisco, CA, USAResearch and Process Development, Sutro Biopharma, Inc, South San Francisco, CA, USAResearch and Process Development, Sutro Biopharma, Inc, South San Francisco, CA, USAResearch and Process Development, Sutro Biopharma, Inc, South San Francisco, CA, USAResearch and Process Development, Sutro Biopharma, Inc, South San Francisco, CA, USAResearch and Process Development, Sutro Biopharma, Inc, South San Francisco, CA, USAResearch and Process Development, Sutro Biopharma, Inc, South San Francisco, CA, USAResearch and Process Development, Sutro Biopharma, Inc, South San Francisco, CA, USATherapeutic bioconjugates are emerging as an essential tool to combat human disease. Site-specific conjugation technologies are widely recognized as the optimal approach for producing homogeneous drug products. Non-natural amino acid (nnAA) incorporation allows the introduction of bioconjugation handles at genetically defined locations. Escherichia coli (E. coli) is a facile host for therapeutic nnAA protein synthesis because it can stably replicate plasmids encoding genes for product and nnAA incorporation. Here, we demonstrate that by engineering E. coli to incorporate high levels of nnAAs, it is feasible to produce nnAA-containing antibody fragments and full-length immunoglobulin Gs (IgGs) in the cytoplasm of E. coli. Using high-density fermentation, it was possible to produce both of these types of molecules with site-specifically incorporated nnAAs at titers > 1 g/L. We anticipate this strategy will help simplify the production and manufacture of promising antibody therapeutics.https://www.tandfonline.com/doi/10.1080/19420862.2024.2316872Antibody–drug conjugatebioconjugatione. coli expressionnon-natural amino acidsite-specifictherapeutic protein |
| spellingShingle | Jacquelyn Blake-Hedges Dan Groff Wilson Foo Jeffrey Hanson Elenor Castillo Miao Wen Diana Cheung Mary Rose Masikat Jian Lu Young Park Nina Abi Carlos Hans Usman Kevin Fong Abigail Yu Sihong Zhou Joyce Kwong Cuong Tran Xiaofan Li Dawei Yuan Trevor Hallam Gang Yin Production of antibodies and antibody fragments containing non-natural amino acids in Escherichia coli mAbs Antibody–drug conjugate bioconjugation e. coli expression non-natural amino acid site-specific therapeutic protein |
| title | Production of antibodies and antibody fragments containing non-natural amino acids in Escherichia coli |
| title_full | Production of antibodies and antibody fragments containing non-natural amino acids in Escherichia coli |
| title_fullStr | Production of antibodies and antibody fragments containing non-natural amino acids in Escherichia coli |
| title_full_unstemmed | Production of antibodies and antibody fragments containing non-natural amino acids in Escherichia coli |
| title_short | Production of antibodies and antibody fragments containing non-natural amino acids in Escherichia coli |
| title_sort | production of antibodies and antibody fragments containing non natural amino acids in escherichia coli |
| topic | Antibody–drug conjugate bioconjugation e. coli expression non-natural amino acid site-specific therapeutic protein |
| url | https://www.tandfonline.com/doi/10.1080/19420862.2024.2316872 |
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