Interplay of heavy chain introns influences efficient transcript splicing and affects product quality of recombinant biotherapeutic antibodies from CHO cells
Introns are included in genes encoding therapeutic proteins for their well-documented function of boosting expression. However, mis-splicing of introns in recombinant immunoglobulin (IgG) heavy chain (HC) transcripts can produce amino acid sequence product variants. These variants can affect product...
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| Language: | English |
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Taylor & Francis Group
2023-12-01
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| Series: | mAbs |
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| Online Access: | https://www.tandfonline.com/doi/10.1080/19420862.2023.2242548 |
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| author | Emma Kelsall Claire Harris Titash Sen Diane Hatton Sarah Dunn Suzanne Gibson |
| author_facet | Emma Kelsall Claire Harris Titash Sen Diane Hatton Sarah Dunn Suzanne Gibson |
| author_sort | Emma Kelsall |
| collection | DOAJ |
| description | Introns are included in genes encoding therapeutic proteins for their well-documented function of boosting expression. However, mis-splicing of introns in recombinant immunoglobulin (IgG) heavy chain (HC) transcripts can produce amino acid sequence product variants. These variants can affect product quality; therefore, purification process optimization may be needed to remove them, or if they cannot be removed, then in-depth characterization must be carried out to understand their effects on biological activity. In this study, HC transgene engineering approaches were investigated and were successful in significantly reducing the previously identified IgG HC splice variants to <0.5%. Subsequently, a comprehensive evaluation was conducted to understand the influence of the different introns in the HC genes on the expression of recombinant biotherapeutic antibodies. The data revealed an unexpected cooperation between specific introns for efficient splicing, where intron retention led to significant reductions in IgG expression of up to 75% for some intron combinations. Furthermore, it was shown that HC introns could be fully removed without significantly affecting productivity. This work paves the way for future biotherapeutic antibody transgene design with regard to inclusion of HC introns. By removing unnecessary introns, transgene mRNA transcript will no longer be mis-spliced, thereby eliminating HC splice variants and improving antibody product quality. |
| format | Article |
| id | doaj-art-ab876963243a4450ba80756faf129466 |
| institution | Kabale University |
| issn | 1942-0862 1942-0870 |
| language | English |
| publishDate | 2023-12-01 |
| publisher | Taylor & Francis Group |
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| series | mAbs |
| spelling | doaj-art-ab876963243a4450ba80756faf1294662025-08-20T03:25:53ZengTaylor & Francis GroupmAbs1942-08621942-08702023-12-0115110.1080/19420862.2023.2242548Interplay of heavy chain introns influences efficient transcript splicing and affects product quality of recombinant biotherapeutic antibodies from CHO cellsEmma Kelsall0Claire Harris1Titash Sen2Diane Hatton3Sarah Dunn4Suzanne Gibson5Cell Culture and Fermentation Sciences, Biopharmaceutical Development, R&D, AstraZeneca, Cambridge, UKCell Culture and Fermentation Sciences, Biopharmaceutical Development, R&D, AstraZeneca, Cambridge, UKCell Culture and Fermentation Sciences, Biopharmaceutical Development, R&D, AstraZeneca, Cambridge, UKCell Culture and Fermentation Sciences, Biopharmaceutical Development, R&D, AstraZeneca, Cambridge, UKCell Culture and Fermentation Sciences, Biopharmaceutical Development, R&D, AstraZeneca, Cambridge, UKCell Culture and Fermentation Sciences, Biopharmaceutical Development, R&D, AstraZeneca, Cambridge, UKIntrons are included in genes encoding therapeutic proteins for their well-documented function of boosting expression. However, mis-splicing of introns in recombinant immunoglobulin (IgG) heavy chain (HC) transcripts can produce amino acid sequence product variants. These variants can affect product quality; therefore, purification process optimization may be needed to remove them, or if they cannot be removed, then in-depth characterization must be carried out to understand their effects on biological activity. In this study, HC transgene engineering approaches were investigated and were successful in significantly reducing the previously identified IgG HC splice variants to <0.5%. Subsequently, a comprehensive evaluation was conducted to understand the influence of the different introns in the HC genes on the expression of recombinant biotherapeutic antibodies. The data revealed an unexpected cooperation between specific introns for efficient splicing, where intron retention led to significant reductions in IgG expression of up to 75% for some intron combinations. Furthermore, it was shown that HC introns could be fully removed without significantly affecting productivity. This work paves the way for future biotherapeutic antibody transgene design with regard to inclusion of HC introns. By removing unnecessary introns, transgene mRNA transcript will no longer be mis-spliced, thereby eliminating HC splice variants and improving antibody product quality.https://www.tandfonline.com/doi/10.1080/19420862.2023.2242548Alternative splicingCHOexpression plasmidintronmonoclonal antibodysplice variant |
| spellingShingle | Emma Kelsall Claire Harris Titash Sen Diane Hatton Sarah Dunn Suzanne Gibson Interplay of heavy chain introns influences efficient transcript splicing and affects product quality of recombinant biotherapeutic antibodies from CHO cells mAbs Alternative splicing CHO expression plasmid intron monoclonal antibody splice variant |
| title | Interplay of heavy chain introns influences efficient transcript splicing and affects product quality of recombinant biotherapeutic antibodies from CHO cells |
| title_full | Interplay of heavy chain introns influences efficient transcript splicing and affects product quality of recombinant biotherapeutic antibodies from CHO cells |
| title_fullStr | Interplay of heavy chain introns influences efficient transcript splicing and affects product quality of recombinant biotherapeutic antibodies from CHO cells |
| title_full_unstemmed | Interplay of heavy chain introns influences efficient transcript splicing and affects product quality of recombinant biotherapeutic antibodies from CHO cells |
| title_short | Interplay of heavy chain introns influences efficient transcript splicing and affects product quality of recombinant biotherapeutic antibodies from CHO cells |
| title_sort | interplay of heavy chain introns influences efficient transcript splicing and affects product quality of recombinant biotherapeutic antibodies from cho cells |
| topic | Alternative splicing CHO expression plasmid intron monoclonal antibody splice variant |
| url | https://www.tandfonline.com/doi/10.1080/19420862.2023.2242548 |
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