Potential of boronic acid functionalized magnetic particles in the adsorption of human antibodies under mammalian cell culture conditions

Borlido, Luis; Azevedo, Ana M.; Roque, Ana C. A.; Aires-Barros, M. Raquel

doi:10.1016/j.chroma.2011.08.084

Cited by 29 publications

(38 citation statements)

References 32 publications

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“…The experimental adsorption values of ligand 22/8 modified MP with dextran and gum arabic coating was found to be 130 and 110 mg hIgG adsorbed g 21 MP, respectively [16]. The commercially available APBA-modified MP shows experimental adsorption of 109 mg hIgG adsorbed g 21 MP [17], whereas MP-BA-3 particles adsorb 173 mg hIgG g 21 MP.…”

Section: Selection Of the Magnetic Materials For Antibody Adsorptionmentioning

confidence: 99%

“…AR is known to be a reporter to study carbohydrate -boronic acid interactions owing to its change in fluorescence intensity upon binding to a boronic acid [20]. MP-BA-3 particles were tested for binding and eluting AR using two types of buffer conditions, previously shown to adsorb and elute IgG from commercial boronic acid particles [17]. From the results shown in figure 2b, it is observed that for binding with phosphate buffer pH 7.4 and elution with glycine buffer pH 11, MP-BA-3 particles adsorbed approximately 40% AR and eluted approximately 30%.…”

Section: Selection Of the Magnetic Materials For Antibody Adsorptionmentioning

confidence: 99%

“…There are protein A-coated MPs commercially available for the purification of antibodies, and previous studies indicated that other types of functionalized MPs can also be suitable for the capture step. In particular, our groups have shown that custom-made MPs coated with small and robust synthetic ligands mimicking protein A [16], and commercial boronic acid-coated MPs [17] represent interesting and viable options for capturing antibodies in purification processes. Commercial boronic acid MPs allowed the direct and selective capture of a human mAb from a CHO feedstock, where binding and elution conditions were tuned to reach maximum yield and final purity [17].…”

Section: Introductionmentioning

confidence: 99%

“…In particular, our groups have shown that custom-made MPs coated with small and robust synthetic ligands mimicking protein A [16], and commercial boronic acid-coated MPs [17] represent interesting and viable options for capturing antibodies in purification processes. Commercial boronic acid MPs allowed the direct and selective capture of a human mAb from a CHO feedstock, where binding and elution conditions were tuned to reach maximum yield and final purity [17]. However, the high cost of these commercial particles makes unviable the scale-up of the purification process.…”

Section: Introductionmentioning

confidence: 99%

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Boronic acid-modified magnetic materials for antibody purification

Dhadge

Hussain

Azevedo

et al. 2014

J. R. Soc. Interface.

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Aminophenyl boronic acids can form reversible covalent ester interactions with cis-diol-containing molecules, serving as a selective tool for binding glycoproteins as antibody molecules that possess oligosaccharides in both the Fv and Fc regions. In this study, amino phenyl boronic acid (APBA) magnetic particles (MPs) were applied for the magnetic separation of antibody molecules. Iron oxide MPs were firstly coated with dextran to avoid non-specific binding and then with 3-glycidyloxypropyl trimethoxysilane to allow further covalent coupling of APBA (APBA_MP MP (Q max). APBA_MPs were also tested for antibody purification directly from CHO cell supernatants. The particles were able to bind 98% of IgG loaded and to recover 95% of pure IgG (purity greater than 98%) at extremely mild conditions.

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Section: Selection Of the Magnetic Materials For Antibody Adsorptionmentioning

confidence: 99%

Section: Selection Of the Magnetic Materials For Antibody Adsorptionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Boronic acid-modified magnetic materials for antibody purification

Dhadge

Hussain

Azevedo

et al. 2014

J. R. Soc. Interface.

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“…Surface functionalization of MNPs with metal ions such as Ni 2+ and Co 2+ has been used to selectively capture polyhistidine-tagged (His-tagged) proteins from cell extracts with subsequent separation using an appropriately applied magnetic field [98–100]. In other studies, boronic acid-functionalized MNPs have been used extensively for one-step capture of human monoclonal antibodies with high yield and purity [101,102]. …”

Section: Magnetic Separationmentioning

confidence: 99%

Bench-To-Bedside Translation of Magnetic Nanoparticles

et al. 2014

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Magnetic nanoparticles (MNPs) are a new and promising addition to the spectrum of biomedicines. Their promise revolves around the broad versatility and biocompatibility of the MNPs and their unique physicochemical properties. Guided by applied external magnetic fields, MNPs represent a cutting-edge tool designed to improve diagnosis and therapy of a broad range of inflammatory, infectious, genetic and degenerative diseases. Magnetic hyperthermia, targeted drug and gene delivery, cell tracking, protein bioseparation and tissue engineering are but a few applications being developed for MNPs. MNPs toxicities linked to shape, size and surface chemistry are real and must be addressed before clinical use is realized. This article presents both the promise and perils of this new nanotechnology, with an eye towards opportunity in translational medical science.

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Stimuli‐Responsive magnetic nanoparticles for monoclonal antibody purification

Borlido

Moura

Azevedo

et al. 2013

Biotechnology Journal

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Monoclonal antibodies (mAbs) are important therapeutic proteins. One of the challenges facing large-scale production of monoclonal antibodies is the capacity bottleneck in downstream processing, which can be circumvented by using magnetic stimuli-responsive polymer nanoparticles. In this work, stimuli-responsive magnetic particles composed of a magnetic poly(methyl methacrylate) core with a poly(N-isopropylacrylamide-co-acrylic acid) (P(NIPAM-co-AA)) shell cross-linked with N, N'-methylenebisacrylamide were prepared by miniemulsion polymerization. The particles were shown to have an average hydrodynamic diameter of 317 nm at 18°C, which decreased to 277 nm at 41°C due to the collapse of the thermo-responsive shell. The particles were superparamagnetic in behavior and exhibited a saturation magnetization of 12.6 emu/g. Subsequently, we evaluated the potential of these negatively charged stimuli-responsive magnetic particles in the purification of a monoclonal antibody from a diafiltered CHO cell culture supernatant by cation exchange. The adsorption of antibodies onto P(NIPAM-co-AA)-coated nanoparticles was highly selective and allowed for the recovery of approximately 94% of the mAb. Different elution strategies were employed providing highly pure mAb fractions with host cell protein (HCP) removal greater than 98%. By exploring the stimuli-responsive properties of the particles, shorter magnetic separation times were possible without significant differences in product yield and purity.

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Potential of boronic acid functionalized magnetic particles in the adsorption of human antibodies under mammalian cell culture conditions

Cited by 29 publications

References 32 publications

Boronic acid-modified magnetic materials for antibody purification

Boronic acid-modified magnetic materials for antibody purification

Bench-To-Bedside Translation of Magnetic Nanoparticles

Stimuli‐Responsive magnetic nanoparticles for monoclonal antibody purification

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