Abstract
Binary mixture adsorption containing Human immunoglobulin (IgG) and Human serum albumin (HSA) on a new cation exchanger mixed mode adsorbent (MabDirect MM) is assessed by batch, fixed bed and expanded bed adsorption experiments. Competitive adsorption isotherm was measured by batch experiment at the binding condition of pH 5.0 without salt where it was found that increasing the IgG concentration present in the solution (0–3 g dm−3) the HSA adsorption capacity decreases significantly (from 29.32 to 3.17 mg gdry−1) as a result of IgG competitive adsorption with HSA on MabDirect MM. Fixed bed adsorption experiments with binary mixture of HSA and IgG were performed at two flowrates. There is no roll up in the adsorption breakthrough curves even when it was increased the experiment timescale (from 2 to 0.7 mL/min at adsorption stage) which suggests that they are both adsorbed also on different sites at the binding condition, 20 mM citrate buffer pH 5.0 with 0.4 M⋅NaCl. Adsorption capacity for both proteins are in agreement with their respective single adsorption isotherms. Displacement experiments were carried out and showed that some amount of protein is released when the other protein is being fed to the column. Expanded bed breakthrough experiment with binary mixture of HSA and IgG shows an adsorption capacity (14.2 mgHSA gdry−1 and 6.3 mgIgG gdry−1) in good agreement with single protein batch experiments and also fixed bed adsorption for similar conditions. Regarding HSA and IgG desorption, it was obtained a HSA recovery of 62% and an IgG recovery of 39%.
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Aboudzadeh, M.R., Aboudzadeh, N., Jiawen, Z., Bin, W.: Binary protein adsorption to DEAE sepharose FF. Korean J. Chem. Eng. 24, 641–647 (2007)
Arévalo, E., Rendueles, M., Fernández, A., Díaz, M.: Equilibrium and simulation of the operation for the adsorption of albumin proteins in an iminodiacetic-Cu bounded ion exchange resin (IMAC). Sep. Purif. Technol. 18, 217–225 (2000)
den-Boer, P., Doeven, M., van der Merwe, J., Lihme, A., Bendix-Hansen, M., Vaarst, I., Pontoppidan, M., Linz, F.: (2013) Rhobust EBA Processing high-cell density mammalian cell cultures: process parameters and performance. DSM Biol.
Gomes, P.F., Loureiro, J.M., Rodrigues, A.E.: Adsorption equilibrium and kinetics of Immunoglobulin G on a mixed-mode adsorbent in batch and packed bed configuration. J. Chromatogr. A 1524, 143–152 (2017a)
Gomes, P.F., Loureiro, J.M., Rodrigues, A.E.: Adsorption of Human Serum Albumin (HSA) on a mixed-mode adsorbent: equilibrium and kinetics. Adsorption 23, 491–505 (2017b)
Gomes, P.F., Loureiro, J.M., Rodrigues, A.E.: Expanded bed adsorption of albumin and immunoglobulin G from human serum onto a cation exchanger mixed mode adsorbent. Adsorption 24, 293–307 (2018)
Kelly, W., Garcia, P., McDermott, S., Mullen, P., Kamguia, G., Jones, G., Ubiera, A., Göklen, K.: Experimental characterization of next-generation expanded-bed adsorbents for capture of a recombinant protein expressed in high-cell-density yeast fermentation. Biotechnol. Appl. Biochem. 60, 510–520 (2013a)
Kelly, W., Kamguia, G., Mullen, P., Ubiera, A., Göklen, K., Huang, Z., Jones, G.: Using a two species competitive binding model to predict expanded bed breakthrough of a recombinant protein expressed in a high cell density fermentation. Biotechnol. Bioprocess Eng. 18, 546–559 (2013b)
Leblebici, P., Leblebici, M.E., Ferreira-da-Silva, F., Rodrigues, A.E., Pais, L.S.: Separation of human immunoglobulin G subclasses on a protein A monolith column. J. Chromatogr. B 962, 89–93 (2014)
Lewus, R.K., Carta, G.: Binary protein adsorption on gel-composite ion-exchange media. Am. Inst. Chem. Eng. J. 45, 512–522 (1999)
Li, P., Xiu, G., Mata, V.G., Grande, C.A., Rodrigues, A.E.: Expanded bed adsorption/desorption of proteins with Streamline Direct CST I adsorbent. Biotechnol. Bioeng. 94, 1155–1163 (2006)
Li, P., Gomes, P.F., Loureiro, J.M., Rodrigues, A.E.: Proteins separation and purification by expanded bed adsorption and simulated moving bed technology. In: Subramanian G (ed.) Continuous processing in pharmaceutical manufacturing, Wiley, Hoboken (2014)
Lima, M.D.A., Freitas, M.D.F.M.D., Gonçalves, L.R.B., Silva Junior, I.Jd: Recovery and purification of a Kluyvermyces lactis β-galactosidase by mixed mode chromatography. J. Chromatogr. B 1015, 181–191 (2016)
Peula, J.M., Hidalgo-Alvarez, R., De Las Nieves, F.J.: Coadsorption of IgG and BSA onto sulfonated polystyrene latex: I. Sequential and competitive coadsorption isotherms. J. Biomater. Sci. Polym. Ed. 7, 231–240 (1996)
Rendueles de la Vega M., Chenou, C., Loureiro, J.M., Rodrigues, A.E.: Mass transfer mechanisms in Hyper D media for chromatographic protein separation. Biochem. Eng. J. 1, 11–23 (1998)
Schaller, J., Gerber, S., Kämpfer, U., Lejon, S., Trachsel, C.: Human blood plasma proteins: structure and function, Wiley, Hoboken, (2008)
Walters, R.R., Graham, J.F., Moore, R.M., Anderson, D.J.: Protein diffusion coefficient measurements by laminar flow analysis: method and applications. Anal. Biochem. 140, 190–195 (1984)
Xia, H.-F., Lin, D.-Q., Yao, S.-J.: Evaluation of new high-density ion exchange adsorbents for expanded bed adsorption chromatography. J. Chromatogr. A 1145, 58–66 (2007)
Xu, X., Lenhoff, A.M.: Binary adsorption of globular proteins on ion-exchange media. J. Chromatogr. A 1216, 6177–6195 (2009)
Yan, J., Zhang, Q.L., Lin, D.Q., Yao, S.J.: Protein adsorption behavior and immunoglobulin separation with a mixed-mode resin based on p-aminohippuric acid. J. Sep. Sci. 37, 2474–2480 (2014)
Yan, J., Zhang, Q.-L., Lin, D.-Q., Yao, S.-J.: Coadsorption of Human Immunoglobulin G and Bovine Serum Albumin on a p-Aminohippuric Acid Based Mixed-Mode Resin. Journal of Chemical & Engineering Data 61, 151–159 (2016)
Yelemane, V., Kangwa, M., Fernández-Lahore, M.: (2016) High-Throughput Screening of Cell Repellent Substrate Chemistry for Application in Expanded Bed Adsorption Chromatography. In: B.D.P, Gummadi SN, Vadlani PV (eds.) Biotechnology and Biochemical Engineering: Select Proceedings of ICACE 2015. Singapore, pp 145–152. https://doi.org/10.1007/978-981-10-1920-3_16
Acknowledgements
The authors would like to thank Upfront Chromatography A/S for kindly providing the adsorbent. This work is a result of project “AIProcMat@N2020 - Advanced Industrial Processes and Materials for a Sustainable Northern Region of Portugal 2020”, with the reference NORTE-01-0145-FEDER-000006, supported by Norte Portugal Regional Operational Programme (NORTE 2020), under the Portugal 2020 Partnership Agreement, through the European Regional Development Fund (ERDF) and of Project POCI-01-0145-FEDER-006984 – Associate Laboratory LSRE-LCM funded by ERDF through COMPETE2020 - Programa Operacional Competitividade e Internacionalização (POCI) – and by national funds through FCT - Fundação para a Ciência e a Tecnologia.
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Gomes, P.F., Loureiro, J.M. & Rodrigues, A.E. Co-adsorption of Albumin and Immunoglobulin G from Human Serum onto a cation exchanger mixed mode adsorbent. Adsorption 24, 745–755 (2018). https://doi.org/10.1007/s10450-018-9984-8
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DOI: https://doi.org/10.1007/s10450-018-9984-8