Non–antigen-contacting region of an asymmetric bispecific antibody to factors IXa/X significantly affects factor VIII-mimetic activity
| Autor: | Atsushi Muto, Takehisa Kitazawa, Kazutaka Yoshihashi, Tetsuo Kojima, Tomoyuki Igawa, Kunihiro Hattori, Miho Funaki, Tetsuhiro Soeda, Satoshi Nakamura, Zenjiro Sampei |
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| Rok vydání: | 2015 |
| Předmět: |
Glycan
constant region Immunology Immunoglobulin light chain Factor IXa IgG subclass Antigen Biomimetic Materials Antibodies Bispecific Humans Immunology and Allergy hinge elbow angle Factor VIII antibody engineering biology Effector Chemistry Fc glycosylation Biological activity Fragment crystallizable region Protein tertiary structure bispecific antibody flexibility Biochemistry Factor X biology.protein Biophysics disulfide bond hemophilia A Antibody Reports |
| Zdroj: | mAbs |
| ISSN: | 1942-0870 1942-0862 |
| DOI: | 10.4161/19420862.2015.989028 |
| Popis: | While antibody engineering improves the properties of therapeutic antibodies, optimization of regions that do not contact antigens has been mainly focused on modifying the effector functions and pharmacokinetics of antibodies. We recently reported an asymmetric anti-FIXa/FX bispecific IgG4 antibody, ACE910, which mimics the cofactor function of FVIII by placing the two factors into spatial proximity for the treatment of hemophilia A. During the optimization process, we found that the activity was significantly affected by IgG subclass and by modifications to the inter-chain disulfide bonds, upper hinge region, elbow hinge region, and Fc glycan, even though these regions were unlikely to come into direct contact with the antigens. Of these non-antigen-contacting regions, the tertiary structure determined by the inter-chain disulfide bonds was found to strongly affect the FVIII-mimetic activity. Interestingly, IgG4-like disulfide bonds between Cys131 in the heavy chain and Cys114 in the light chain, and disulfide bonds between the two heavy chains at the hinge region were indispensable for the high FVIII-mimetic activity. Moreover, proline mutations in the upper hinge region and removal of the Fc glycan enhanced the FVIII-mimetic activity, suggesting that flexibility of the upper hinge region and the Fc portion structure are important for the FVIII-mimetic activity. This study suggests that these non-antigen-contacting regions can be engineered to improve the biological activity of IgG antibodies with functions similar to ACE910, such as placing two antigens into spatial proximity, retargeting effector cells to target cells, or co-ligating two identical or different antigens on the same cell. |
| Databáze: | OpenAIRE |
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