Purification of human antibodies from animal cell cultures using gum arabic coated magnetic particles

Abstract

BACKGROUND The emergence of monoclonal antibodies (mAbs) as new biopharmaceutical products requires the development of new purification methods that are not only effective but are able to reduce production costs. To address the problematic recovery of mAbs, gum arabic (GA) coated magnetic particles (MPs) were used for the purification of human antibodies from animal cells supernatants. RESULTS MPs were synthesized via co‐precipitation and exhibited a spherical‐like physical aspect, with an average hydrodynamic diameter of 473 nm and a zeta potential of –26 mV. The adsorption and elution of IgG on these adsorbents was thoroughly studied. Adsorption of human IgG was enhanced at pH 6, for which a qmax of 244 mg IgG g−1 MPs and Kd of 25 mg L−1 were obtained. Increasing salt concentrations at a basic pH (1 mol L−1 NaCl at pH 11) were found to improve the elution of bound IgG. The MPs were challenged with an artificial protein mixture containing human IgG, albumin, insulin and apo‐transferrin. An overall yield of 84% was achieved, retrieving 92% of bound IgG. CONCLUSIONS MPs were successfully used for the capture of monoclonal antibodies from two distinct mammalian cell cultures, a Chinese hamster ovary (CHO) and a hybridoma cell culture supernatants. The elution yields were high, ranging between 84% and 94%, with overall yields ranging from 72% to 88%. Final purities of 85% were reached for hybridoma cell supernatants. © 2014 Society of Chemical Industry

Polydopamine assisted preparation of Ti4+‐decorated magnetic particles for selective and rapid adsorption of phosphorylated proteins

“…Over the past decade, magnetic particles (MPs) have gained rapid and substantial progress, and have had a significant impact on protein adsorption . MPs‐based methods are time‐effectiveness and labor‐saving since MPs as adsorbents could be conveniently separated by applying an external magnetic field and no centrifugation step is needed to realize the phase separation.…”

Recovery and Purification of (Bio)Pharmaceuticals Using (Nano)Materials

“…In addition, the recovery of both proteins was above 95%, with a purity of 95% and 81% for Lys and IgG, respectively [121]. Alves et al [122] synthetized and tested MPs of iron oxide coated with gum arabic for the purification of antibodies. It was proved that using these low-cost adsorbents it was possible to purify IgG from the supernatant mammalian cell cultures, namely Chinese hamster ovary (CHO) and Hybridoma, by a cation exchange mechanism, achieving elution yields of 93.5%.…”

“…Given the simplicity and reduced costs of this purification strategy, the results obtained are very encouraging. In addition, the possibility of reusing MPs is an obvious economic advantage [122]. The same iron oxide, coated with gum arabic magnetic supports, were functionalized with two different triazine-based affinity ligands -ligand 22/8 (artificial Protein A) and ligand 8/7 (artificial Protein L) -with high specificity towards IgG [59].…”

Direct Affinity Ligand Immobilization onto Bare Iron Oxide Nanoparticles Enables Efficient Magnetic Separation of Antibodies

“…We consider magnetic separation a highly promising alternative, which has found increasing popularity in recent years. − ,− This technique is based on nonporous magnetic particles that are freely dispersed in the process fluid and can be controlled magnetically. The nonporous magnetic particle adsorbent leads to reduced mass transfer limitations compared to conventional porous chromatography beds, which favors fast target adsorption/desorption and, thus, high throughput. , Furthermore, clogging of the purification matrix is prevented due to the nonporosity, enabling the direct processing of unclarified cell culture broth and process intensification. , Another powerful characteristic of the technique is the scale-independence of the magnetic adsorption process, which simplifies the scale-up …”