دورية أكاديمية
Effective flow-through polishing strategies for knob-into-hole bispecific antibodies.
| العنوان: | Effective flow-through polishing strategies for knob-into-hole bispecific antibodies. |
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| المؤلفون: | Chen SW; Downstream Processing Group, Bioprocessing Technology Institute, Agency for Science, Technology and Research, Singapore, Singapore., Hoi KM; Downstream Processing Group, Bioprocessing Technology Institute, Agency for Science, Technology and Research, Singapore, Singapore., Mahfut FB; Cell Line Development Group, Bioprocessing Technology Institute, Agency for Science, Technology and Research, Singapore, Singapore., Yang Y; Cell Line Development Group, Bioprocessing Technology Institute, Agency for Science, Technology and Research, Singapore, Singapore., Zhang W; Downstream Processing Group, Bioprocessing Technology Institute, Agency for Science, Technology and Research, Singapore, Singapore. zhang_wei@bti.a-star.edu.sg. |
| المصدر: | Bioresources and bioprocessing [Bioresour Bioprocess] 2022 Sep 14; Vol. 9 (1), pp. 98. Date of Electronic Publication: 2022 Sep 14. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Springer-Verlag, GmbH Country of Publication: Germany NLM ID: 101665551 Publication Model: Electronic Cited Medium: Internet ISSN: 2197-4365 (Electronic) Linking ISSN: 21974365 NLM ISO Abbreviation: Bioresour Bioprocess Subsets: PubMed not MEDLINE |
| أسماء مطبوعة: | Original Publication: Heidelberg, Germany : Springer-Verlag, GmbH, [2014]- |
| مستخلص: | Bispecific antibodies (bsAbs), though possessing great therapeutic potential, are extremely challenging to obtain at high purity within a limited number of scalable downstream processing steps. Complementary to Protein A chromatography, polishing strategies play a critical role at removing the remaining high molecular weight (HMW) and low molecular weight (LMW) species, as well as host cell proteins (HCP) in order to achieve a final product of high purity. Here, we demonstrate using two knob-into-hole (KiH) bsAb constructs that two flow-through polishing steps utilising Capto Butyl ImpRes and Capto adhere resins, performed after an optimal Protein A affinity chromatography step can further reduce the HCP by 17- to 35-fold as well as HMW and LMW species with respect to monomer by ~ 4-6% and ~ 1%, respectively, to meet therapeutical requirement at 30-60 mg/mL-resin (R) load. This complete flow-through polishing strategy, guided by Design of Experiments (DoE), eliminates undesirable aggregation problems associated with the higher aggregation propensity of scFv containing bsAbs that may occur in the bind and elute mode, offering an improved ease of overall process operation without additional elution buffer preparation and consumption, thus aligning well with process intensification efforts. Overall, we demonstrate that through the employment of (1) Protein A chromatography step and (2) flow-through polishing steps, a final product containing < 1% HMW species, < 1% LMW species and < 100 ppm HCP can be obtained with an overall process recovery of 56-87%. (© 2022. The Author(s).) |
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| فهرسة مساهمة: | Keywords: Bispecific antibody; Flow-through polishing; High molecular weight impurities; Higher aggregation propensity; Host cell proteins; Knob-into-hole; Low molecular weight impurities |
| تواريخ الأحداث: | Date Created: 20240422 Latest Revision: 20240425 |
| رمز التحديث: | 20240425 |
| مُعرف محوري في PubMed: | PMC10992779 |
| DOI: | 10.1186/s40643-022-00590-8 |
| PMID: | 38647877 |
| قاعدة البيانات: | MEDLINE |
Find this article in full text from Springer Verlag. 
Full Text Finder | تدمد: | 2197-4365 |
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| DOI: | 10.1186/s40643-022-00590-8 |