التفاصيل البيبلوغرافية
| العنوان: |
Development of a platform process for the production and purification of single‐domain antibodies |
| المؤلفون: |
Crowell, Laura E, Goodwine, Chaz, Holt, Carla S, Rocha, Lucia, Vega, Celina, Rodriguez, Sergio A, Dalvie, Neil C, Tracey, Mary K, Puntel, Mariana, Wigdorovitz, Andrés, Parreño, Viviana, Love, Kerry R, Cramer, Steven M, Love, J Christopher |
| المساهمون: |
Koch Institute for Integrative Cancer Research at MIT, Massachusetts Institute of Technology. Department of Chemical Engineering, Massachusetts Institute of Technology. Department of Biological Engineering |
| المصدر: |
Wiley |
| بيانات النشر: |
Wiley |
| سنة النشر: |
2021 |
| المجموعة: |
DSpace@MIT (Massachusetts Institute of Technology) |
| الوصف: |
Single-domain antibodies (sdAbs) offer the affinity and therapeutic value of conventional antibodies, with increased stability and solubility. Unlike conventional antibodies, however, sdAbs do not benefit from a platform manufacturing process. While successful production of a variety of sdAbs has been shown in numerous hosts, purification methods are often molecule specific or require affinity tags, which generally cannot be used in clinical manufacturing due to regulatory concerns. Here, we have developed a broadly applicable production and purification process for sdAbs in Komagataella phaffii (Pichia pastoris) and demonstrated the production of eight different sdAbs at a quality appropriate for nonclinical studies. We developed a two-step, integrated purification process without the use of affinity resins and showed that modification of a single process parameter, pH of the bridging buffer, was required for the successful purification of a variety of sdAbs. Further, we determined that this parameter can be predicted based only on the biophysical characteristics of the target molecule. Using these methods, we produced nonclinical quality sdAbs as few as 5 weeks after identifying the product sequence. Nonclinical studies of three different sdAbs showed that molecules produced using our platform process conferred protection against viral shedding of rotavirus or H1N1 influenza and were equivalent to similar molecules produced in Escherichia coli and purified using affinity tags. |
| نوع الوثيقة: |
article in journal/newspaper |
| وصف الملف: |
application/pdf |
| اللغة: |
English |
| العلاقة: |
Biotechnology and Bioengineering; https://hdl.handle.net/1721.1/135524Test |
| الإتاحة: |
https://hdl.handle.net/1721.1/135524Test |
| حقوق: |
Creative Commons Attribution 4.0 International license ; http://creativecommons.org/licenses/by-nc-nd/4.0Test/ |
| رقم الانضمام: |
edsbas.3104731D |
| قاعدة البيانات: |
BASE |
