Discovery and investigation of the truncation of the (GGGGS)n linker and its effect on the productivity of bispecific antibodies expressed in mammalian cells.
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| Title: | Discovery and investigation of the truncation of the (GGGGS)n linker and its effect on the productivity of bispecific antibodies expressed in mammalian cells. |
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| Authors: | Fang, Yan1 (AUTHOR) yan1.fang@qilu-pharma.com, Chen, Xi1 (AUTHOR), Sun, Zhen1 (AUTHOR), Yan, Xiaodan1 (AUTHOR), Shi, Lani1 (AUTHOR), Jin, Congcong1 (AUTHOR) |
| Source: | Bioprocess & Biosystems Engineering. Jan2025, Vol. 48 Issue 1, p159-170. 12p. |
| Subjects: | Chimeric proteins, Protein engineering, Therapeutic use of proteins, Immune response, Proteolytic enzymes |
| Abstract: | Protein engineering is a powerful tool for designing or modifying therapeutic proteins for enhanced efficacy, increased safety, reduced immunogenicity, and improved delivery. Fusion proteins are an important group of therapeutic compounds that often require an ideal linker to combine diverse domains to fulfill the desired function. GGGGS [(G4S)n] linkers are commonly used during the engineering of proteins because of their flexibility and resistance to proteases. However, unexpected truncation was observed in the linker of a bispecific antibody, which presented challenges in terms of production and quality. In this work, a bispecific antibody containing 5*G4S was investigated, and the truncation position of the linkers was confirmed. Our investigation revealed that codon optimization, which can overcome the negative influence of a high repetition rate and high GC content in the (G4S)n linker, may reduce the truncation rate from 5–10% to 1–5%. Moreover, the probability of truncation when a shortened 3* or 4*G4S linker was used was much lower than that when a 5*G4S linker was used in mammalian cells. In the case of expressing a bispecific antibody, the bioactivity and purity of the product containing a shorter G4S linker were further investigated and are discussed. [ABSTRACT FROM AUTHOR] |
| Copyright of Bioprocess & Biosystems Engineering is the property of Springer Nature and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.) | |
| Database: | Engineering Source |
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| Header | DbId: egs DbLabel: Engineering Source An: 182471291 AccessLevel: 6 PubType: Academic Journal PubTypeId: academicJournal PreciseRelevancyScore: 0 |
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| Items | – Name: Title Label: Title Group: Ti Data: Discovery and investigation of the truncation of the (GGGGS)n linker and its effect on the productivity of bispecific antibodies expressed in mammalian cells. – Name: Author Label: Authors Group: Au Data: <searchLink fieldCode="AR" term="%22Fang%2C+Yan%22">Fang, Yan</searchLink><relatesTo>1</relatesTo> (AUTHOR)<i> yan1.fang@qilu-pharma.com</i><br /><searchLink fieldCode="AR" term="%22Chen%2C+Xi%22">Chen, Xi</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Sun%2C+Zhen%22">Sun, Zhen</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Yan%2C+Xiaodan%22">Yan, Xiaodan</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Shi%2C+Lani%22">Shi, Lani</searchLink><relatesTo>1</relatesTo> (AUTHOR)<br /><searchLink fieldCode="AR" term="%22Jin%2C+Congcong%22">Jin, Congcong</searchLink><relatesTo>1</relatesTo> (AUTHOR) – Name: TitleSource Label: Source Group: Src Data: <searchLink fieldCode="JN" term="%22Bioprocess+%26+Biosystems+Engineering%22">Bioprocess & Biosystems Engineering</searchLink>. Jan2025, Vol. 48 Issue 1, p159-170. 12p. – Name: Subject Label: Subjects Group: Su Data: <searchLink fieldCode="DE" term="%22Chimeric+proteins%22">Chimeric proteins</searchLink><br /><searchLink fieldCode="DE" term="%22Protein+engineering%22">Protein engineering</searchLink><br /><searchLink fieldCode="DE" term="%22Therapeutic+use+of+proteins%22">Therapeutic use of proteins</searchLink><br /><searchLink fieldCode="DE" term="%22Immune+response%22">Immune response</searchLink><br /><searchLink fieldCode="DE" term="%22Proteolytic+enzymes%22">Proteolytic enzymes</searchLink> – Name: Abstract Label: Abstract Group: Ab Data: Protein engineering is a powerful tool for designing or modifying therapeutic proteins for enhanced efficacy, increased safety, reduced immunogenicity, and improved delivery. Fusion proteins are an important group of therapeutic compounds that often require an ideal linker to combine diverse domains to fulfill the desired function. GGGGS [(G4S)n] linkers are commonly used during the engineering of proteins because of their flexibility and resistance to proteases. However, unexpected truncation was observed in the linker of a bispecific antibody, which presented challenges in terms of production and quality. In this work, a bispecific antibody containing 5*G4S was investigated, and the truncation position of the linkers was confirmed. Our investigation revealed that codon optimization, which can overcome the negative influence of a high repetition rate and high GC content in the (G4S)n linker, may reduce the truncation rate from 5–10% to 1–5%. Moreover, the probability of truncation when a shortened 3* or 4*G4S linker was used was much lower than that when a 5*G4S linker was used in mammalian cells. In the case of expressing a bispecific antibody, the bioactivity and purity of the product containing a shorter G4S linker were further investigated and are discussed. [ABSTRACT FROM AUTHOR] – Name: AbstractSuppliedCopyright Label: Group: Ab Data: <i>Copyright of Bioprocess & Biosystems Engineering is the property of Springer Nature and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.) |
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| RecordInfo | BibRecord: BibEntity: Identifiers: – Type: doi Value: 10.1007/s00449-024-03100-6 Languages: – Code: eng Text: English PhysicalDescription: Pagination: PageCount: 12 StartPage: 159 Subjects: – SubjectFull: Chimeric proteins Type: general – SubjectFull: Protein engineering Type: general – SubjectFull: Therapeutic use of proteins Type: general – SubjectFull: Immune response Type: general – SubjectFull: Proteolytic enzymes Type: general Titles: – TitleFull: Discovery and investigation of the truncation of the (GGGGS)n linker and its effect on the productivity of bispecific antibodies expressed in mammalian cells. Type: main BibRelationships: HasContributorRelationships: – PersonEntity: Name: NameFull: Fang, Yan – PersonEntity: Name: NameFull: Chen, Xi – PersonEntity: Name: NameFull: Sun, Zhen – PersonEntity: Name: NameFull: Yan, Xiaodan – PersonEntity: Name: NameFull: Shi, Lani – PersonEntity: Name: NameFull: Jin, Congcong IsPartOfRelationships: – BibEntity: Dates: – D: 01 M: 01 Text: Jan2025 Type: published Y: 2025 Identifiers: – Type: issn-print Value: 16157591 Numbering: – Type: volume Value: 48 – Type: issue Value: 1 Titles: – TitleFull: Bioprocess & Biosystems Engineering Type: main |
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