Affinity ATPS Strategies for the Selective Fractionation of Biomolecules

Ruiz‐Ruiz, Federico; Benavides, Jorge; Rito‐Palomares, Marco

doi:10.1007/978-3-319-59309-8_6

Cited by 4 publications

(2 citation statements)

References 58 publications

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“… 77 Affinity ligands may also be incorporated into ATPE to improve the selectivity of the process. 78 Such an approach at preparative scale can boast a high ligand concentration per volume of the polymer solution, allowing for higher productivity. 79 Although numerous applications of ATPE have been demonstrated over several decades, 80 a continuous pilot 81 and mini‐plant‐scale 82 ATPE system for mAb purification was demonstrated more recently.…”

Section: Continuous Downstream Processesmentioning

confidence: 99%

Developments and opportunities in continuous biopharmaceutical manufacturing

Khanal

Lenhoff

2021

mAbs

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Today’s biologics manufacturing practices incur high costs to the drug makers, which can contribute to high prices for patients. Timely investment in the development and implementation of continuous biomanufacturing can increase the production of consistent-quality drugs at a lower cost and a faster pace, to meet growing demand. Efficient use of equipment, manufacturing footprint, and labor also offer the potential to improve drug accessibility. Although technological efforts enabling continuous biomanufacturing have commenced, challenges remain in the integration, monitoring, and control of traditionally segmented unit operations. Here, we discuss recent developments supporting the implementation of continuous biomanufacturing, along with their benefits.

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Section: Continuous Downstream Processesmentioning

confidence: 99%

Developments and opportunities in continuous biopharmaceutical manufacturing

Khanal

Lenhoff

2021

mAbs

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“…Although these systems are usually less selective when compared with the conventional methods, their selectivity may be improved by introducing a specific ligand to steer the biomolecule of interest into one of the phases [22]. In this approach, called affinity partition, the presence of affinity ligands, i.e., molecules with specificity and biorecognition properties towards a target solute, allows for the extraction of the target from a crude feedstock despite possible similarities with the contaminants [22].…”

Section: Introductionmentioning

confidence: 99%

Initial Screening of Poly(ethylene glycol) Amino Ligands for Affinity Purification of Plasmid DNA in Aqueous Two-Phase Systems

Silva

Jorge

Martins

et al. 2021

Life

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Gene therapy and DNA vaccination are among the most expected biotechnological and medical advances for the coming years. However, the lack of cost-effective large-scale production and purification of pharmaceutical-grade plasmid DNA (pDNA) still hampers their wide application. Downstream processing, which is mainly chromatography-based, of pDNA remains the key manufacturing step. Despite its high resolution, the scaling-up of chromatography is usually difficult and presents low capacity, resulting in low yields. Alternative methods that are based on aqueous two-phase systems (ATPSs) have been studied. Although higher yields may be obtained, its selectivity is often low. In this work, modified polymers based on poly(ethylene glycol) (PEG) derivatisation with amino groups (PEG–amine) or conjugation with positively charged amino acids (PEG–lysine, PEG–arginine, and PEG–histidine) were studied to increase the selectivity of PEG–dextran systems towards the partition of a model plasmid. A two-step strategy was employed to obtain suitable pure formulations of pDNA. In the first step, a PEG–dextran system with the addition of the affinity ligand was used with the recovery of the pDNA in the PEG-rich phase. Then, the pDNA was re-extracted to an ammonium-sulphate-rich phase in the second step. After removing the salt, this method yielded a purified preparation of pDNA without RNA and protein contamination.

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