Large-Scale Protein Chromatography

Bertolini, Joseph; Gomme, Peter T.; Thomas, Patrick E.

doi:10.1385/1-59259-742-4:211

Cited by 3 publications

(3 citation statements)

References 2 publications

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“…After the optional addition of charged groups ( 5), a fraction of remaining hydroxyl groups was esterified with 2-chloropro- pionoyl chloride to produce the ATRP initiator-functionalized surface (6). When using limiting quantities of 2-chlorpropionoyl chloride, incorporation of ATRP initiators via esterification had low reaction efficiency (13%), perhaps due to small amounts of water remaining on the base material.…”

Section: Aqueous Atrp Graft Polymerization Of Pdma Frommentioning

confidence: 99%

“…Therefore, there is considerable interest in new strategies for synthesizing SEC media that permit one to carefully “tune” the properties of the stationary phase to maximum separation efficiency over a desired molecular weight range. This need has intensified in recent years due to the growth of proteomics, which requires highly efficient strategies for separating complex mixtures, and by the shift in bioprocessing of blood products away from the classic cold ethanol precipitation process of Cohn to much more efficient and selective chromatography trains. , Human blood plasma is a source material for many life-saving therapeutics, including albumin, intravenous immune globulin, α-antiplasmin, α 1 -antitrypsin, and a number of coagulation proteins. As it is now thought to contain more than 1500 unique peptides, , human plasma has the potential to yield a number of new protein-based therapeutics.…”

Section: Introductionmentioning

confidence: 99%

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Synthesis of Novel Size Exclusion Chromatography Support by Surface Initiated Aqueous Atom Transfer Radical Polymerization

et al. 2007

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We report the use of aqueous surface-initiated atom transfer radical polymerization (SI-ATRP) to grow polymer brushes from a "gigaporous" polymeric chromatography support for use as a novel size exclusion chromatography medium. Poly(N,N-dimethylacrylamide) (PDMA) was grown from hydrolyzable surface initiators via SI-ATRP catalyzed by 1,1,4,7,10,10-hexamethyltriethylenetetramine (HMTETA)/CuCl. Grafted polymer was characterized semiquantitatively by ATR-FTIR and also cleaved and quantitatively characterized for mass, molecular weight, and polydispersity via analytical SEC/MALLS. The synthesis provides control over graft density and allows the creation of dense brushes. Incorporation of negative surface charge was found to be crucial for improving the initiation efficiency. As polymer molecular weight and density could be controlled through reaction conditions, the resulting low-polydispersity grafted polymer brush medium is shown to be suitable for use as a customizable size exclusion chromatography medium for investigating the principals of entropic interaction chromatography. All packed media investigated showed size-dependent partitioning of solutes, even for low graft density systems. Increasing the molecular weight of the grafts allowed solutes more access to the volume fraction in the column available for partitioning. Compared to low graft density media, increased graft density caused eluted solute probes to be retained less within the column and allowed for greater size discrimination of probes whose molecular weights were less than 10(4) kDa.

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Section: Aqueous Atrp Graft Polymerization Of Pdma Frommentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Synthesis of Novel Size Exclusion Chromatography Support by Surface Initiated Aqueous Atom Transfer Radical Polymerization

et al. 2007

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“…Since then, it has been used at preparative scales in the manufacture of insulin (Janson, 1971) and in the purification of human serum albumin (HSA) from blood plasma (Lin et al, 2000;Curling, 1980). First reported by Friedli and Kistler (1972), the removal of ethanol from HSA purified by the Cohn-Oncley cold-ethanol fractionation process remains one of the largest and most well-documented applications of SEC (Johnston and Adcock, 2000;Bertolini et al, 2004).…”

Section: Introductionmentioning

confidence: 99%

Entropic interaction chromatography: Separating proteins on the basis of size using end‐grafted polymer brushes

Pang

Koska

Coad

et al. 2005

Biotech & Bioengineering

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Partitioning of a macromolecule into the interfacial volume occupied by a grafted polymer brush decreases the configurational entropy (ÁS c brush ) of the terminally attached linear polymer chains due to a loss of free volume. Self-consistent field theory (SCF) calculations are used to show that ÁS c brush is a strong function of both the size (MW p ) of the partitioning macromolecule and the depth of penetration into the brush volume. We further demonstrate that the strong dependence of ÁS c brush on MW p provides a novel and powerful platform, which we call entropic interaction chromatography (EIC), for efficiently separating mixtures of proteins on the basis of size. Two EIC columns, differing primarily in polymer grafting density, were prepared by growing a brush of poly(methoxyethyl acrylamide) chains on the surface of a widepore (1,000-Å pores, 64-Am diameter rigid beads) resin (Toyopearl AF-650M) bearing surface aldehyde groups. Semipreparative 0.1-L columns packed with either EIC resin provide reduced-plate heights of 2 or less for efficient separation of globular protein mixtures over at least three molecular-weight decades. Protein partitioning within these wide-pore EIC columns is shown to be effectively modeled as a thermodynamically controlled process, allowing partition coefficients (K P ) and elution chromatograms to be accurately predicted using a column model that combines SCF calculation of K P values with an equilibrium-dispersion type model of solute transport through the column. This model is used to explore the dependence of column separation efficiency on brush properties, predicting that optimal separation of proteins over a broad MW p range is achieved at low to moderate grafting densities and intermediate chain lengths. B 2005 Wiley Periodicals, Inc.

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Functional monolithic platforms: Chromatographic tools for antibody purification

Barroso

Hussain

Roque

et al. 2013

Biotechnology Journal

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Polymer monoliths are an efficient platform for antibody purification. The use of monoclonal antibodies (mAbs) and engineered antibody structures as therapeutics has increased exponentially over the past few decades. Several approaches use polymer monoliths to purify large quantities of antibody with defined clinical and performance requirements. Functional monolithic supports have attracted a great deal of attention as they offer practical advantages for antibody purification, such as more rapid analysis, smaller sample volume requirements and the opportunity for a greater target molecule enrichment. This review focuses on the development of synthetic and natural polymer-based monoliths for antibody purification. The materials and methods employed in monolith production are discussed, highlighting the properties of each system. We also review the structural characterization techniques available using monolithic systems and their performance under different chromatographic approaches to antibody capture and release. Finally, a summary of monolithic platforms developed for antibody separation is presented, as well as expected trends in research to solve current and future challenges in this field. This review comprises a comprehensive analysis of proposed solutions highlighting the remarkable potential of monolithic platforms.

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Large-Scale Protein Chromatography

Cited by 3 publications

References 2 publications

Synthesis of Novel Size Exclusion Chromatography Support by Surface Initiated Aqueous Atom Transfer Radical Polymerization

Synthesis of Novel Size Exclusion Chromatography Support by Surface Initiated Aqueous Atom Transfer Radical Polymerization

Entropic interaction chromatography: Separating proteins on the basis of size using end‐grafted polymer brushes

Functional monolithic platforms: Chromatographic tools for antibody purification

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