IgG Aggregation Mechanism for CHO Cell Lines Expressing Excess Heavy Chains
Aggregates in protein therapeutics like IgG monoclonal antibodies (mAb) are detrimental to product safety and efficacy. It has been reported that aggregates form in Chinese hamster ovary (CHO) cell lines expressing greater amount of heavy chain (HC) than light chain (LC). In this study, we observed...
| Main Authors: | , , , |
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| Format: | Journal Article |
| Language: | English |
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2016
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| Online Access: | https://hdl.handle.net/10356/81469 http://hdl.handle.net/10220/40791 |
| _version_ | 1811693635859644416 |
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| author | Ho, Steven C. L. Wang, Tianhua Song, Zhiwei Yang, Yuansheng |
| author2 | School of Chemical and Biomedical Engineering |
| author_facet | School of Chemical and Biomedical Engineering Ho, Steven C. L. Wang, Tianhua Song, Zhiwei Yang, Yuansheng |
| author_sort | Ho, Steven C. L. |
| collection | NTU |
| description | Aggregates in protein therapeutics like IgG monoclonal antibodies (mAb) are detrimental to product safety and efficacy. It has been reported that aggregates form in Chinese hamster ovary (CHO) cell lines expressing greater amount of heavy chain (HC) than light chain (LC). In this study, we observed that aggregates could form within the cells with excess HC and were partially secreted into the supernatant. The aggregates in the supernatant consisted of mainly HC and were partially dissociated under either reducing or denaturing conditions. Mutation of a predicted free cysteine on HC to prevent disulfide bonding did not reduce aggregation. Re-transfecting CHO cells with excess HC with more BiP, an important IgG molecular chaperone, partially reduced unwanted aggregates and fragments possibly by helping retain more incomplete products within the cell for either proper assembly or degradation. A second transfection of LC into CHO cells with excess HC to increase the LC expression to a level greater than the HC expression successfully removed all aggregates and fragments. mAb product aggregation in CHO cells with excess HC occur due to a combination of limited chaperones and LC:HC ratio. These results provide added insights to aggregate formation and would be useful for development of mAb cell lines with reduced aggregates. |
| first_indexed | 2024-10-01T06:54:49Z |
| format | Journal Article |
| id | ntu-10356/81469 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T06:54:49Z |
| publishDate | 2016 |
| record_format | dspace |
| spelling | ntu-10356/814692023-12-29T06:51:25Z IgG Aggregation Mechanism for CHO Cell Lines Expressing Excess Heavy Chains Ho, Steven C. L. Wang, Tianhua Song, Zhiwei Yang, Yuansheng School of Chemical and Biomedical Engineering Monoclonal antibody Chinese hamster ovary cells BiP IgG aggregation Aggregates in protein therapeutics like IgG monoclonal antibodies (mAb) are detrimental to product safety and efficacy. It has been reported that aggregates form in Chinese hamster ovary (CHO) cell lines expressing greater amount of heavy chain (HC) than light chain (LC). In this study, we observed that aggregates could form within the cells with excess HC and were partially secreted into the supernatant. The aggregates in the supernatant consisted of mainly HC and were partially dissociated under either reducing or denaturing conditions. Mutation of a predicted free cysteine on HC to prevent disulfide bonding did not reduce aggregation. Re-transfecting CHO cells with excess HC with more BiP, an important IgG molecular chaperone, partially reduced unwanted aggregates and fragments possibly by helping retain more incomplete products within the cell for either proper assembly or degradation. A second transfection of LC into CHO cells with excess HC to increase the LC expression to a level greater than the HC expression successfully removed all aggregates and fragments. mAb product aggregation in CHO cells with excess HC occur due to a combination of limited chaperones and LC:HC ratio. These results provide added insights to aggregate formation and would be useful for development of mAb cell lines with reduced aggregates. ASTAR (Agency for Sci., Tech. and Research, S’pore) Accepted version 2016-06-24T04:46:56Z 2019-12-06T14:31:44Z 2016-06-24T04:46:56Z 2019-12-06T14:31:44Z 2015 Journal Article Ho, S. C. L., Wang, T., Song, Z., & Yang, Y. (2015). IgG Aggregation Mechanism for CHO Cell Lines Expressing Excess Heavy Chains. Molecular Biotechnology, 57(7), 625-634. 1073-6085 https://hdl.handle.net/10356/81469 http://hdl.handle.net/10220/40791 10.1007/s12033-015-9852-7 en Molecular Biotechnology © 2015 Springer Science+Business Media New York. This is the author created version of a work that has been peer reviewed and accepted for publication by Molecular Biotechnology, Springer Science+Business Media New York. It incorporates referee’s comments but changes resulting from the publishing process, such as copyediting, structural formatting, may not be reflected in this document. The published version is available at: [http://dx.doi.org/10.1007/s12033-015-9852-7]. 30 p. application/pdf |
| spellingShingle | Monoclonal antibody Chinese hamster ovary cells BiP IgG aggregation Ho, Steven C. L. Wang, Tianhua Song, Zhiwei Yang, Yuansheng IgG Aggregation Mechanism for CHO Cell Lines Expressing Excess Heavy Chains |
| title | IgG Aggregation Mechanism for CHO Cell Lines Expressing Excess Heavy Chains |
| title_full | IgG Aggregation Mechanism for CHO Cell Lines Expressing Excess Heavy Chains |
| title_fullStr | IgG Aggregation Mechanism for CHO Cell Lines Expressing Excess Heavy Chains |
| title_full_unstemmed | IgG Aggregation Mechanism for CHO Cell Lines Expressing Excess Heavy Chains |
| title_short | IgG Aggregation Mechanism for CHO Cell Lines Expressing Excess Heavy Chains |
| title_sort | igg aggregation mechanism for cho cell lines expressing excess heavy chains |
| topic | Monoclonal antibody Chinese hamster ovary cells BiP IgG aggregation |
| url | https://hdl.handle.net/10356/81469 http://hdl.handle.net/10220/40791 |
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