Simultaneous Maturation of Single Chain Antibody Stability and Affinity by CHO Cell Display
Antibody stability and affinity are two important features of its applications in therapy and diagnosis. Antibody display technologies such as yeast and bacterial displays have been successfully used for improving both affinity and stability. Although mammalian cell display has also been utilized fo...
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MDPI AG
2022-08-01
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| Series: | Bioengineering |
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| Online Access: | https://www.mdpi.com/2306-5354/9/8/360 |
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| author | Ruiqi Luo Baole Qu Lili An Yun Zhao Yang Cao Peng Ren Haiying Hang |
| author_facet | Ruiqi Luo Baole Qu Lili An Yun Zhao Yang Cao Peng Ren Haiying Hang |
| author_sort | Ruiqi Luo |
| collection | DOAJ |
| description | Antibody stability and affinity are two important features of its applications in therapy and diagnosis. Antibody display technologies such as yeast and bacterial displays have been successfully used for improving both affinity and stability. Although mammalian cell display has also been utilized for maturing antibody affinity, it has not been applied for improving antibody stability. Previously, we developed a Chinese hamster ovary (CHO) cell display platform in which activation-induced cytidine deaminase (AID) was used to induce antibody mutation, and antibody affinity was successfully matured using the platform. In the current study, we developed thermo-resistant (TR) CHO cells for the purpose of maturing both antibody stability and affinity. We cultured TR CHO cells displaying an antibody mutant library and labeled them at temperatures above 41 °C, enriching cells that displayed antibody mutants with both the highest affinities and the highest display levels. To evaluate our system, we chose three antibodies to improve their affinities and stabilities. We succeeded in simultaneously improving both affinities and stabilities of all three antibodies. Of note, we obtained an anti-TNFα antibody mutant with a Tm (dissolution temperature) value 12 °C higher and affinity 160-fold greater than the parent antibody after two rounds of cell proliferation and flow cytometric sorting. By using CHO cells with its advantages in protein folding, post-translational modifications, and code usage, this procedure is likely to be widely used in maturing antibodies and other proteins in the future. |
| first_indexed | 2024-03-09T10:01:27Z |
| format | Article |
| id | doaj.art-58fbe95307264ec7b3ecaee7eacbc280 |
| institution | Directory Open Access Journal |
| issn | 2306-5354 |
| language | English |
| last_indexed | 2024-03-09T10:01:27Z |
| publishDate | 2022-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Bioengineering |
| spelling | doaj.art-58fbe95307264ec7b3ecaee7eacbc2802023-12-01T23:25:02ZengMDPI AGBioengineering2306-53542022-08-019836010.3390/bioengineering9080360Simultaneous Maturation of Single Chain Antibody Stability and Affinity by CHO Cell DisplayRuiqi Luo0Baole Qu1Lili An2Yun Zhao3Yang Cao4Peng Ren5Haiying Hang6Key Laboratory for Protein and Peptide Pharmaceuticals, National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, ChinaKey Laboratory for Protein and Peptide Pharmaceuticals, National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, ChinaKey Laboratory for Protein and Peptide Pharmaceuticals, National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, ChinaKey Laboratory for Protein and Peptide Pharmaceuticals, National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, ChinaCenter of Growth, Metabolism and Aging, Key Laboratory of Bio-Resources and Eco-Environment of Ministry of Education, College of Life Sciences, Sichuan University, Chengdu 610064, ChinaDepartment of Thoracic Surgery, Peking University Third Hospital, Beijing 100191, ChinaKey Laboratory for Protein and Peptide Pharmaceuticals, National Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, ChinaAntibody stability and affinity are two important features of its applications in therapy and diagnosis. Antibody display technologies such as yeast and bacterial displays have been successfully used for improving both affinity and stability. Although mammalian cell display has also been utilized for maturing antibody affinity, it has not been applied for improving antibody stability. Previously, we developed a Chinese hamster ovary (CHO) cell display platform in which activation-induced cytidine deaminase (AID) was used to induce antibody mutation, and antibody affinity was successfully matured using the platform. In the current study, we developed thermo-resistant (TR) CHO cells for the purpose of maturing both antibody stability and affinity. We cultured TR CHO cells displaying an antibody mutant library and labeled them at temperatures above 41 °C, enriching cells that displayed antibody mutants with both the highest affinities and the highest display levels. To evaluate our system, we chose three antibodies to improve their affinities and stabilities. We succeeded in simultaneously improving both affinities and stabilities of all three antibodies. Of note, we obtained an anti-TNFα antibody mutant with a Tm (dissolution temperature) value 12 °C higher and affinity 160-fold greater than the parent antibody after two rounds of cell proliferation and flow cytometric sorting. By using CHO cells with its advantages in protein folding, post-translational modifications, and code usage, this procedure is likely to be widely used in maturing antibodies and other proteins in the future.https://www.mdpi.com/2306-5354/9/8/360stabilityaffinity maturationmammalian cell displaythermo-resistant CHO cells |
| spellingShingle | Ruiqi Luo Baole Qu Lili An Yun Zhao Yang Cao Peng Ren Haiying Hang Simultaneous Maturation of Single Chain Antibody Stability and Affinity by CHO Cell Display Bioengineering stability affinity maturation mammalian cell display thermo-resistant CHO cells |
| title | Simultaneous Maturation of Single Chain Antibody Stability and Affinity by CHO Cell Display |
| title_full | Simultaneous Maturation of Single Chain Antibody Stability and Affinity by CHO Cell Display |
| title_fullStr | Simultaneous Maturation of Single Chain Antibody Stability and Affinity by CHO Cell Display |
| title_full_unstemmed | Simultaneous Maturation of Single Chain Antibody Stability and Affinity by CHO Cell Display |
| title_short | Simultaneous Maturation of Single Chain Antibody Stability and Affinity by CHO Cell Display |
| title_sort | simultaneous maturation of single chain antibody stability and affinity by cho cell display |
| topic | stability affinity maturation mammalian cell display thermo-resistant CHO cells |
| url | https://www.mdpi.com/2306-5354/9/8/360 |
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