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Engineering an anti-CD52 antibody for enhanced deamidation stability
| Content Provider | Scilit |
|---|---|
| Author | Qiu, Huawei Wei, Ronnie Jaworski, Julie Boudanova, Ekaterina Hughes, Heather Vanpatten, Scott Lund, Anders Day, Jaime Zhou, Yanfeng McSherry, Tracey Pan, Clark Q. Sendak, Rebecca |
| Copyright Year | 2019 |
| Description | Journal: mAbs Deamidation evaluation and mitigation is an important aspect of therapeutic antibody developability assessment. We investigated the structure and function of the Asn-Gly deamidation in a human anti-CD52 IgG1 antibody light chain complementarity-determining region 1, and risk mitigation through protein engineering. Antigen binding affinity was found to decrease about 400-fold when $Asn^{33}$ was replaced with an Asp residue to mimic the deamidation product, suggesting significant impacts on antibody function. Other variants made at $Asn^{33}$ (N33H, N33Q, N33H, N33R) were also found to result in significant loss of antigen binding affinity. The co-crystal structure of the antigen-binding fragment bound to a CD52 peptide mimetic was solved at 2.2Å (PDB code 6OBD), which revealed that $Asn^{33}$ directly interacts with the CD52 phosphate group via a hydrogen bond. $Gly^{34}$, but sits away from the binding interface, rendering it more amendable to mutagenesis without affecting affinity. Saturation mutants at $Gly^{34}$ were prepared and subjected to forced deamidation by incubation at elevated pH and temperature. Three mutants (G34R, G34K and G34Q) showed increased resistance to deamidation by LC-MS peptide mapping, while maintaining high binding affinity to CD52 antigen measured by Biacore. A complement -dependent cytotoxicity assay indicated that these mutants function by triggering antibody effector function. This study illustrates the importance of structure-based design and extensive mutagenesis to mitigate antibody developability issues. |
| Related Links | https://www.tandfonline.com/doi/pdf/10.1080/19420862.2019.1631117?needAccess=true https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6748592/pdf |
| Ending Page | 1275 |
| Page Count | 10 |
| Starting Page | 1266 |
| ISSN | 19420862 |
| e-ISSN | 19420870 |
| DOI | 10.1080/19420862.2019.1631117 |
| Journal | mAbs |
| Issue Number | 7 |
| Volume Number | 11 |
| Language | English |
| Publisher | Informa UK Limited |
| Publisher Date | 2019-10-03 |
| Access Restriction | Open |
| Subject Keyword | Journal: mAbs Biochemistry and Molecular Biology Antibody Engineering Developability Structure-function |
| Content Type | Text |
| Subject | Immunology and Allergy Immunology |
