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Shokri, Atefeh
Publications (2 of 2) Show all publications
Brechmann, N. A., Eriksson, P.-O., Eriksson, K., Oscarsson, S., Buijs, J., Shokri, A., . . . Chotteau, V. (2019). Pilot-scale process for magnetic bead purification of antibodies directly from non-clarified CHO cell culture. Biotechnology progress (Print)
Open this publication in new window or tab >>Pilot-scale process for magnetic bead purification of antibodies directly from non-clarified CHO cell culture
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2019 (English)In: Biotechnology progress (Print), ISSN 8756-7938, E-ISSN 1520-6033Article in journal (Refereed) Published
Abstract [en]

High capacity magnetic protein A agarose beads, LOABeads PrtA, were used in the development

of a new process for affinity purification of monoclonal antibodies (mAbs) from non-clarified

CHO cell broth using a pilot-scale magnetic separator. The LOABeads had a maximum binding

capacity of 65 mg/mL and an adsorption capacity of 25–42 mg IgG/mL bead in suspension for an

IgG concentration of 1 to 8 g/L. Pilot-scale separation was initially tested in a mAb capture step

from 26 L clarified harvest. Small-scale experiments showed that similar mAb adsorptions were

obtained in cell broth containing 40 Å~ 106 cells/mL as in clarified supernatant. Two pilot-scale

purification runs were then performed on non-clarified cell broth from fed-batch runs of 16 L,

where a rapid mAb adsorption ≥96.6% was observed after 1 h. This process using 1 L of magnetic beads had an overall mAb yield of 86% and 16 times concentration factor. After this single protein

A capture step, the mAb purity was similar to the one obtained by column chromatography, while

the host cell protein content was very low, <10 ppm. Our results showed that this magnetic bead

mAb purification process, using a dedicated pilot-scale separation device, was a highly efficient

single step, which directly connected the culture to the downstream process without cell clarification.

Purification of mAb directly from non-clarified cell broth without cell separation can provide

significant savings in terms of resources, operation time, and equipment, compared to legacy procedure of cell separation followed by column chromatography step.

Place, publisher, year, edition, pages
AIChE, 2019
Keywords
magnetic beads, purification, monoclonal antibody, pilot-scale, downstream-bioprocess
National Category
Bioprocess Technology
Research subject
Biotechnology
Identifiers
urn:nbn:se:kth:diva-248987 (URN)10.1002/btpr.2775 (DOI)2-s2.0-85061063787 (Scopus ID)
Projects
AdBIOPRO
Funder
Vinnova, 2016-04152Vinnova, 2016-05181
Note

QC 20190429

Available from: 2019-04-11 Created: 2019-04-11 Last updated: 2019-04-29Bibliographically approved
Al-Khalili, L., Gillner, K., Zhang, Y., Åstrand, C., Shokri, A., Hughes-Brittain, N., . . . Chotteau, V. (2016). Characterization of Human CD133+Cells in Biocompatible Poly(l-lactic acid) Electrospun Nano-Fiber Scaffolds. Journal of Biomaterials and Tissue Engineering, 6(12), 959-966
Open this publication in new window or tab >>Characterization of Human CD133+Cells in Biocompatible Poly(l-lactic acid) Electrospun Nano-Fiber Scaffolds
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2016 (English)In: Journal of Biomaterials and Tissue Engineering, ISSN 2157-9083, E-ISSN 2157-9091, Vol. 6, no 12, p. 959-966Article in journal (Refereed) Published
Abstract [en]

CD133+ cells are potential myogenic progenitors for skeletal muscle regeneration to treat muscular dystrophies. The proliferation of human CD133+ stem cells was studied for 14 days in 3D biomimetic electrospun poly-L-lactic acid (PLLA) nano-fiber scaffolds. Additionally, the myogenic differentiation of the cells was studied during the last 7 days of the culture period. The cells were homogeneously distributed in the 3D scaffolds while colony formation and myotube formation occurred in 2D. After a lag phase due to lower initial cell attachment and an adaptation period, the cell growth rate in 3D was comparable to 2D after 7 and 14 days of culture. The expression of the stem cell (SC) marker PAX7 was 1.5-fold higher in 3D than 2D while the differentiation markers MyoG, Desmin and MyoD were only slightly changed (or remain unchanged) in 3D but strongly increased in 2D (12.6, 3.9, and 7.9-fold), and the myotube formation observed in 2D was absent in 3D. The marker expression during proliferation and differentiation, together with the absence of myotubes in 3D, indicates a better maintenance of stemness in 3D PLLA and stronger tendency for spontaneous differentiation in 2D culture. This makes 3D PLLA a promising biomaterial for the expansion of functional CD133+ cells.

Place, publisher, year, edition, pages
American Scientific Publishers, 2016
Keywords
Myogenic Progenitor Cell, CD133+Cells, Myogenic Differentiation, 3D Cell Culturing, Electrospun Biodegradable Nano-Fiber Scaffold
National Category
Biological Sciences
Identifiers
urn:nbn:se:kth:diva-198952 (URN)10.1166/jbt.2016.1531 (DOI)000387148500005 ()2-s2.0-84998636398 (Scopus ID)
Funder
EU, FP7, Seventh Framework Programme
Note

QC 20170113

Available from: 2017-01-13 Created: 2016-12-22 Last updated: 2017-11-29Bibliographically approved
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