TY - JOUR
T1 - Fishing human monoclonal antibodies from a CHO cell supernatant with boronic acid magnetic particles
AU - Borlido, Luis
AU - Azevedo, Ana M.
AU - Sousa, A. G.
AU - Oliveira, P. H.
AU - Roque, Ana Cecília Afonso
AU - Aires-Barros, Maria Raquel
N1 - Sem PDF.
L. Borlido acknowledges Fundacao para a Ciencia e Tecnologia (FCT) for the Ph.D. fellowship BD/45077/2008. A. M. Azevedo acknowledges the initiative "Ciencia 2007" of the Portuguese Ministry for Science, Technology, and Higher Education (http://www.mctes.pt/). The authors would also like to acknowledge FCT for the financial support through the contract PTDC/EBB-BIO/098961/2008.
PY - 2012
Y1 - 2012
N2 - In this work we have evaluated the potential of boronic acid functionalized magnetic particles for the one-step capture of a human monoclonal antibody (mAb) from a Chinese hamster ovary (CHO) cell culture supernatant. For comparison, Protein A coated magnetic particles were also used. The most important factor influencing the overall process yield and product purity in boronic acid particles was found to be the binding pH. Basic pH values promoted higher purities while resulting in decreased yields due to the competing effects of molecules such as glucose and lactate present in the cell culture supernatant. After optimization, the particles were successfully used in a multi-cycle purification process of the mAb from the CHO feedstock. Boronic acid particles were able to achieve an average overall yield of 86% with 88% removal of CHO host cell proteins (HCP) when the binding was performed at pH 7.4, while at pH 8.5 these values were 58% and 97%, respectively. In both cases, genomic DNA removal was in excess of 97%. Comparatively, Protein A particles recorded an average overall yield of 80% and an HCP removal greater than 99%. The adsorption of the mAb to the boronic acid particles was shown to be mediated by strong affinity interactions. Overall, boronic acid based purification processes can offer a cost-effective alternative to Protein A as the direct capturing step from the mammalian cell culture.
AB - In this work we have evaluated the potential of boronic acid functionalized magnetic particles for the one-step capture of a human monoclonal antibody (mAb) from a Chinese hamster ovary (CHO) cell culture supernatant. For comparison, Protein A coated magnetic particles were also used. The most important factor influencing the overall process yield and product purity in boronic acid particles was found to be the binding pH. Basic pH values promoted higher purities while resulting in decreased yields due to the competing effects of molecules such as glucose and lactate present in the cell culture supernatant. After optimization, the particles were successfully used in a multi-cycle purification process of the mAb from the CHO feedstock. Boronic acid particles were able to achieve an average overall yield of 86% with 88% removal of CHO host cell proteins (HCP) when the binding was performed at pH 7.4, while at pH 8.5 these values were 58% and 97%, respectively. In both cases, genomic DNA removal was in excess of 97%. Comparatively, Protein A particles recorded an average overall yield of 80% and an HCP removal greater than 99%. The adsorption of the mAb to the boronic acid particles was shown to be mediated by strong affinity interactions. Overall, boronic acid based purification processes can offer a cost-effective alternative to Protein A as the direct capturing step from the mammalian cell culture.
KW - Magnetic particles
KW - Boronic acid
KW - Monoclonal antibody
KW - Affinity purification;
KW - Downstream processing
U2 - 10.1016/j.jchromb.2012.07.014
DO - 10.1016/j.jchromb.2012.07.014
M3 - Article
C2 - 22857861
VL - 903
SP - 163
EP - 170
JO - Journal of chromatography. B, Analytical technologies in the biomedical and life sciences
JF - Journal of chromatography. B, Analytical technologies in the biomedical and life sciences
SN - 1873-376X
ER -