التفاصيل البيبلوغرافية
| العنوان: |
Biophysical characterization and stability of modified IgG1 antibodies with different hexamerization propensities |
| المؤلفون: |
van Kampen, Muriel D, Kuipers-De Wilt, Leonie H A M, van Egmond, Mariëlle L, Reinders-Blankert, Petra, van den Bremer, Ewald T J, Wang, Guanbo, Heck, Albert J R, Parren, Paul W H I, Beurskens, Frank J, Schuurman, Janine, de Jong, Rob N |
| المساهمون: |
Sub Biomol.Mass Spectrometry & Proteom., Afd Biomol.Mass Spect. and Proteomics, Dep Scheikunde, Biomolecular Mass Spectrometry and Proteomics |
| سنة النشر: |
2022 |
| مصطلحات موضوعية: |
Biophysical properties, Conformational and Colloidal stability, Developability, HexaBody, Hexamerization, Reversible self-association, Pharmaceutical Science |
| الوصف: |
The hexamerization of natural, human IgG antibodies after cell surface antigen binding can induce activation of the classical complement pathway. Mutations stimulating Fc domain-mediated hexamerization can potentiate complement activation and induce the clustering of cell surface receptors, a finding that was applied to different clinically investigated antibody therapeutics. Here, we biophysically characterized how increased self-association of IgG1 antibody variants with different hexamerization propensity may impact their developability, rather than functional properties. Self-Interaction Chromatography, Dynamic Light Scattering and PEG-induced precipitation showed that IgG variant self-association at neutral pH increased in the order wild type (WT) < E430G < E345K < E345R < E430G-E345R-S440Y, consistent with functional activity. Self-association was strongly pH-dependent, and single point mutants were fully monomeric at pH 5. Differential Scanning Calorimetry and Fluorimetry showed that mutation E430G decreased conformational stability. Interestingly, heat-induced unfolding facilitated by mutation E430G was reversible at 60°C, while a solvent-exposed hydrophobic mutation caused irreversible aggregation. Remarkably, neither increased dynamic self-association propensity at neutral pH nor decreased conformational stability substantially affected the stability of concentrated variants E430G or E345K during storage for two years at 2-8°C. We discuss how these findings may inform the design and development of IgG-based therapeutics. |
| نوع الوثيقة: |
article in journal/newspaper |
| وصف الملف: |
application/pdf |
| اللغة: |
English |
| تدمد: |
0022-3549 |
| Relation: |
https://dspace.library.uu.nl/handle/1874/421385 |
| الاتاحة: |
https://dspace.library.uu.nl/handle/1874/421385 |
| Rights: |
info:eu-repo/semantics/OpenAccess |
| رقم الانضمام: |
edsbas.41C098B9 |
| قاعدة البيانات: |
BASE |