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  • 1.
    Al-Khalili, Lubna
    et al.
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Gillner, Karin
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Zhang, Ye
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Åstrand, Carolina
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Shokri, Atefeh
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Hughes-Brittain, Nanaaya
    McKean, Robert
    Robb, Brendan
    Chotteau, Véronique
    KTH, Skolan för bioteknologi (BIO), Industriell bioteknologi.
    Characterization of Human CD133+Cells in Biocompatible Poly(l-lactic acid) Electrospun Nano-Fiber Scaffolds2016Ingår i: Journal of Biomaterials and Tissue Engineering, ISSN 2157-9083, E-ISSN 2157-9091, Vol. 6, nr 12, s. 959-966Artikel i tidskrift (Refereegranskat)
    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.

  • 2.
    Brechmann, Nils Arnold
    et al.
    KTH, Skolan för kemi, bioteknologi och hälsa (CBH), Industriell bioteknologi. AdBIOPRO, VINNOVA Competence Centre for Advanced BioProduction by Continuous Processing, Stockholm, Sweden.
    Eriksson, Per-Olov
    Eriksson, Kristofer
    Oscarsson, Sven
    Buijs, Jos
    Shokri, Atefeh
    KTH, Skolan för kemi, bioteknologi och hälsa (CBH), Industriell bioteknologi. AdBIOPRO, VINNOVA Competence Centre for Advanced BioProduction by Continuous Processing, Stockholm, Sweden.
    Hjälm, Göran
    Chotteau, Véronique
    KTH, Skolan för kemi, bioteknologi och hälsa (CBH), Industriell bioteknologi. AdBIOPRO, VINNOVA Competence Centre for Advanced BioProduction by Continuous Processing, Stockholm, Sweden.
    Pilot-scale process for magnetic bead purification of antibodies directly from non-clarified CHO cell culture2019Ingår i: Biotechnology progress (Print), ISSN 8756-7938, E-ISSN 1520-6033Artikel i tidskrift (Refereegranskat)
    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.

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