Clarification of a monoclonal antibody with cationic polyelectrolytes: Analysis of influencing parameters

Sieberz, Julia; Çınar, Elif; Wohlgemuth, Kerstin; Schembecker, Gerhard

doi:10.1016/j.bej.2017.02.011

Cited by 3 publications

(2 citation statements)

References 45 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…While in contrast, polymers with primary amines complex with BSA strongly, leading to higher turbidity T max and precipitate or coexisted solid–liquid coacervate complexes (Figure d). Similar results have been founded in previous studies, where primary amines typically endow strong complexation and solid-like coacervates. , These findings validate that the spacer and charge groups, as well as the structure of polyelectrolytes, all impact the charge interaction and coacervate complexes with BSA protein.…”

Section: Resultssupporting

confidence: 90%

Effects of Control Factors on Protein–Polyelectrolyte Complex Coacervation

Zhou,

Wan,

Cohen Stuart

et al. 2023

Biomacromolecules

View full text Add to dashboard Cite

Protein−polyelectrolyte complex coacervation is of particular interest for mimicking intracellular phase separation and organization. Yet, the challenge arises from regulating the coacervation due to the globular structure and anisotropic distributed charges of protein. Herein, we fully investigate the different control factors and reveal their effects on proteinpolyelectrolyte coacervation. We prepared mixtures of BSA (bovine serum albumin) with different cationic polymers, which include linear and branched polyelectrolytes covering different spacer and charge groups, chain lengths, and polymer structures. With BSA-PDMAEMA [poly(N,N-dimethylaminomethyl methacrylate)] as the main investigated pair, we find that the moderate pH and ionic strength are essential for the adequate electrostatic interaction and formation of coacervate droplets. For most BSA−polymer mixtures, excess polyelectrolytes are required to achieve the full complexation, as evidenced by the deviated optimal charge mixing ratios from the charge stoichiometry. Polymers with longer chains or primary amine groups and a branched structure endow a strong electrostatic interaction with BSA and cause a bigger charge ratio deviation associated with the formation of solid-like coacervate complexes. Nevertheless, both the liquid-and solid-like coacervates hardly interrupt the BSA structure and activity, indicating the safe encapsulation of proteins by the coacervation with polyelectrolytes. Our study validates the crucial control of the diverse factors in regulating protein−polyelectrolyte coacervation, and the revealed principles shall be instructive for establishing other protein-based coacervations and boosting their potential applications.

show abstract

Section: Resultssupporting

confidence: 90%

Effects of Control Factors on Protein–Polyelectrolyte Complex Coacervation

Zhou,

Wan,

Cohen Stuart

et al. 2023

Biomacromolecules

View full text Add to dashboard Cite

show abstract

“…Solms et al developed a thermodynamic framework for the description of a phase diagram of a lysozyme–ovalbumin system [ 25 ]. Sieberz et al investigated the influence of model proteins as contaminants on the precipitation behavior of mAbs during an polyelectrolyte preciptiation process [ 26 ]. All these findings suggest, on the one hand, that the protein–protein interaction influences the protein phase behavior and, on the other, that this influence is not simply a linear or otherwise geometric combination of pure species interaction terms [ 27 ].…”

Section: Introductionmentioning

confidence: 99%

Precipitation of complex antibody solutions: influence of contaminant composition and cell culture medium on the precipitation behavior

Großhans

Suhm

Hubbuch

2019

Bioprocess Biosyst Eng

View full text Add to dashboard Cite

Preparative protein precipitation is known as a cost-efficient and easy-to-use alternative to chromatographic purification steps. This said, at the moment, there is no process for monoclonal antibodies (mAb) on the market, although especially polyethylene glycol-induced precipitation has shown great potential. One reason might be the highly complex behavior of each component of a crude feedstock during the precipitation process. For different investigated mAbs, significant variations in the host cell protein (HCP) reduction are observed. In contrast to the precipitation behavior of single components, the interactions and interplay in a complex feedstock are not fully understood yet. This work discusses the influence of contaminants on the precipitation behavior of two different mAbs, an IgG1, and an IgG2. By spiking the mAbs with mock solution, a complex feedstock could successfully be mimicked. Spiking contaminants influenced the yield and purity of the mAbs after the precipitation step, compared to the precipitation behavior of the single components. The mixture showed a decrease in the contaminant and mAb solubility. By re-buffering the mock solution prior to spiking, special salts, small molecules like amino acids, vitamins, or sugars could be depleted while larger ones like HCP or DNA were still present. Therefore, it was possible to distinguish the influence of small molecules and larger ones. Hence, mAb–macromolecular interaction could be identified as a possible reason for the observed higher precipitation propensity, while small molecules of the cell culture medium were identified as solubilisation factors during the precipitation process.

show abstract

Emerging biomaterials for downstream manufacturing of therapeutic proteins

Stern

Lock

et al. 2019

Acta Biomaterialia

View full text Add to dashboard Cite

Clarification of a monoclonal antibody with cationic polyelectrolytes: Analysis of influencing parameters

Cited by 3 publications

References 45 publications

Effects of Control Factors on Protein–Polyelectrolyte Complex Coacervation

Effects of Control Factors on Protein–Polyelectrolyte Complex Coacervation

Precipitation of complex antibody solutions: influence of contaminant composition and cell culture medium on the precipitation behavior

Emerging biomaterials for downstream manufacturing of therapeutic proteins

Contact Info

Product

Resources

About