The Humanization and Maturation of an Anti-PrPc Antibody
The cellular prion protein (PrPc) is a cell surface glycoprotein that is highly expressed in a variety of cancer tissues in addition to the nervous system, and its elevated expression is correlated to poor prognosis in many cancer patients. Our team previously found that patients with colorectal can...
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MDPI AG
2024-02-01
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| Series: | Bioengineering |
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| Online Access: | https://www.mdpi.com/2306-5354/11/3/242 |
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| author | Cheng Zhang Fanlei Ran Lei Du Xiaohui Wang Lei Liu Jinming Liu Quan Chen Yang Cao Lijun Bi Haiying Hang |
| author_facet | Cheng Zhang Fanlei Ran Lei Du Xiaohui Wang Lei Liu Jinming Liu Quan Chen Yang Cao Lijun Bi Haiying Hang |
| author_sort | Cheng Zhang |
| collection | DOAJ |
| description | The cellular prion protein (PrPc) is a cell surface glycoprotein that is highly expressed in a variety of cancer tissues in addition to the nervous system, and its elevated expression is correlated to poor prognosis in many cancer patients. Our team previously found that patients with colorectal cancer (CRC) with high-level PrPc expression had significantly poorer survival than those with no or low-level PrPc expression. Mouse antibodies for PrPc inhibited tumor initiation and liver metastasis of PrPc-positive human CRC cells in mouse model experiments. PrPc is a candidate target for CRC therapy. In this study, we newly cloned a mouse anti-PrPc antibody (Clone 6) and humanized it, then affinity-matured this antibody using a CHO cell display with a peptide antigen and full-length PrPc, respectively. We obtained two humanized antibody clones with affinities toward a full-length PrPc of about 10- and 100-fold of that of the original antibody. The two humanized antibodies bound to the PrPc displayed significantly better on the cell surface than Clone 6. Used for Western blotting and immunohistochemistry, the humanized antibody with the highest affinity is superior to the two most frequently used commercial antibodies (8H4 and 3F4). The two new antibodies have the potential to be developed as useful reagents for PrPc detection and even therapeutic antibodies targeting PrPc-positive cancers. |
| first_indexed | 2024-04-24T18:32:04Z |
| format | Article |
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| issn | 2306-5354 |
| language | English |
| last_indexed | 2024-04-24T18:32:04Z |
| publishDate | 2024-02-01 |
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| series | Bioengineering |
| spelling | doaj.art-2d5f41fc159748389014095198ab7e0e2024-03-27T13:21:51ZengMDPI AGBioengineering2306-53542024-02-0111324210.3390/bioengineering11030242The Humanization and Maturation of an Anti-PrPc AntibodyCheng Zhang0Fanlei Ran1Lei Du2Xiaohui Wang3Lei Liu4Jinming Liu5Quan Chen6Yang Cao7Lijun Bi8Haiying Hang9Key Laboratory of RNA Biology, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, ChinaKey Laboratory of RNA Biology, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, ChinaThe State Key Laboratory of Membrane Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, ChinaThe State Key Laboratory of Membrane Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, ChinaThe State Key Laboratory of Membrane Biology, Institute of Zoology, Chinese Academy of Sciences, Beijing 100101, ChinaThe State Key Laboratory of Medicinal Chemical Biology, College of Life Sciences, Nankai University, Tianjin 300071, ChinaThe State Key Laboratory of Medicinal Chemical Biology, College of Life Sciences, Nankai University, Tianjin 300071, 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, ChinaKey Laboratory of RNA Biology, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, ChinaKey Laboratory of RNA Biology, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, ChinaThe cellular prion protein (PrPc) is a cell surface glycoprotein that is highly expressed in a variety of cancer tissues in addition to the nervous system, and its elevated expression is correlated to poor prognosis in many cancer patients. Our team previously found that patients with colorectal cancer (CRC) with high-level PrPc expression had significantly poorer survival than those with no or low-level PrPc expression. Mouse antibodies for PrPc inhibited tumor initiation and liver metastasis of PrPc-positive human CRC cells in mouse model experiments. PrPc is a candidate target for CRC therapy. In this study, we newly cloned a mouse anti-PrPc antibody (Clone 6) and humanized it, then affinity-matured this antibody using a CHO cell display with a peptide antigen and full-length PrPc, respectively. We obtained two humanized antibody clones with affinities toward a full-length PrPc of about 10- and 100-fold of that of the original antibody. The two humanized antibodies bound to the PrPc displayed significantly better on the cell surface than Clone 6. Used for Western blotting and immunohistochemistry, the humanized antibody with the highest affinity is superior to the two most frequently used commercial antibodies (8H4 and 3F4). The two new antibodies have the potential to be developed as useful reagents for PrPc detection and even therapeutic antibodies targeting PrPc-positive cancers.https://www.mdpi.com/2306-5354/11/3/242anti-PrPc antibodyaffinity maturationmammalian cell displayhumanizationcolorectal cancer |
| spellingShingle | Cheng Zhang Fanlei Ran Lei Du Xiaohui Wang Lei Liu Jinming Liu Quan Chen Yang Cao Lijun Bi Haiying Hang The Humanization and Maturation of an Anti-PrPc Antibody Bioengineering anti-PrPc antibody affinity maturation mammalian cell display humanization colorectal cancer |
| title | The Humanization and Maturation of an Anti-PrPc Antibody |
| title_full | The Humanization and Maturation of an Anti-PrPc Antibody |
| title_fullStr | The Humanization and Maturation of an Anti-PrPc Antibody |
| title_full_unstemmed | The Humanization and Maturation of an Anti-PrPc Antibody |
| title_short | The Humanization and Maturation of an Anti-PrPc Antibody |
| title_sort | humanization and maturation of an anti prpc antibody |
| topic | anti-PrPc antibody affinity maturation mammalian cell display humanization colorectal cancer |
| url | https://www.mdpi.com/2306-5354/11/3/242 |
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