Direct chromatographic capture of enzyme from crude homogenate using immobilized metal affinity chromatography on a continuous supermacroporous adsorbent

Arvidsson, Per-Ola; Plieva, Fatima M.; Lozinsky, V. I.; Galaev, Igor Yu.; Mattìasson, Bo

doi:10.1016/s0021-9673(02)01871-x

Cited by 245 publications

(185 citation statements)

References 11 publications

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“…Monolithic macroporous polyacrylamide gels, socalled cryogels, have recently been developed for applications in bioseparation (25)(26)(27)(28) and are characterized by high porosity and elasticity. Because of the size (10-100 m) and interconnected structure of the pores and the absence of nonspecific interactions with the adsorbent, cells pass freely through native cryogels without affinity ligands (28).…”

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confidence: 99%

“…2B). It has been demonstrated by scanning electron-microscopy studies that cells captured by affinity cryogel monoliths are bound to the plain ''flat'' parts of the pore walls and are not entrapped in ''dead flow'' zones (25,32). Possible reasons for the disruption of affinity bonds may be the deformation of the surface and͞or the damage to affinity ligands caused by mechanical compression.…”

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confidence: 99%

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Detachment of affinity-captured bioparticles by elastic deformation of a macroporous hydrogel

Dainiak

Kumar

Galaev

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

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Adsorption of bioparticles to affinity surfaces involves polyvalent interactions, complicating greatly the recovery of the adsorbed material. A unique system for the efficient binding and release of different cells and particles is described. Affinity-bound bioparticles and synthetic particles are detached from the macroporous hydrogel matrix, a so-called cryogel, when the cryogel undergoes elastic deformation. The particle detachment upon elastic deformation is believed to be due to breaking of many of the multipoint attachments between the particles and the affinity matrix and the change in the distance between affinity ligands when the matrix is deformed. However, no release of affinity-bound protein occurred upon elastic deformation. The phenomenon of particle detachment upon elastic deformation is believed to be of a generic nature, because it was demonstrated for a variety of bioparticles of different sizes and for synthetic particles, for different ligandreceptor pairs (IgG-protein A, sugar-ConA, metal ion-chelating ligand), and when the deformation was caused by either external forces (mechanical deformation) or internal forces (the shrinkage of thermosensitive, macroporous hydrogel upon an increase in temperature). The elasticity of cryogel monoliths ensures high recovery of captured cells under mild conditions, with highly retained viability. This property, along with their continuous porous structure makes cryogel monoliths very attractive for applications in affinity cell separation. affinity cryogel monoliths ͉ polyvalent interactions ͉ cell separation ͉ thermosensitive hydrogels ͉ cell release

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confidence: 99%

mentioning

confidence: 99%

Detachment of affinity-captured bioparticles by elastic deformation of a macroporous hydrogel

Dainiak

Kumar

Galaev

et al. 2006

Proc. Natl. Acad. Sci. U.S.A.

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“…Cryogel monolith preparation Epoxy-activated monoliths were produced as previously described [37]. In brief, epoxy-activated monoliths (plugs 12.5 x 7.1 mm) were produced using 3.5% solution of co-monomers (Acrylamide, N,N`methylenebis(acrylamide), and allyl glycidyl ether) in the reaction mixture with ammonium persulfate and TEMED as initiators.…”

Section: Methodsmentioning

confidence: 99%

“…In brief, epoxy-activated monoliths (plugs 12.5 x 7.1 mm) were produced using 3.5% solution of co-monomers (Acrylamide, N,N`methylenebis(acrylamide), and allyl glycidyl ether) in the reaction mixture with ammonium persulfate and TEMED as initiators. Coupling IDA ligands to epoxy activated monoliths was carried out according to [37]. The amount of chelate groups was determined to be 13 µmol/ml adsorbent by assaying the amount of bound Cu2+ ions.…”

Section: Methodsmentioning

confidence: 99%

“…These properties make monolithic adsorbents highly suitable for the processing of particulate-containing samples. Affinity monoliths have previously been exploited for the fractionation of different types of mammalian [34] and bacterial [35] cells, affinity capture of viruses [36] and for integrated capture/purification of recombinant proteins from non-clarified cell culture fluids [37,38]. Chelating affinity monoliths with immobilised Cu2+ will bind mitochondria, most likely through histidineresidues on their surface.…”

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Binding mitochondria to cryogel monoliths allows detection of proteins specifically released following permeability transition

Teilum

Hansson

Dainiak

et al. 2006

Analytical Biochemistry

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Monolithic High‐Performance Liquid Chromatography Columns

Desire

Hilder

Arrua

2017

Encyclopedia of Analytical Chemistry

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Monolithic HPLC (high‐performance liquid chromatography) columns have emerged over the recent decades as highly attractive alternatives to conventional HPLC technology. A monolith is considered to be a continuous piece of porous material. Two types of monolithic columns have been developed for liquid chromatography (LC) applications; they are polymer monoliths and silica monoliths, which differ based on their porous properties. Polymer monoliths consist solely of macropores (pores larger than 50 nm), while silica monoliths possess both macropores and mesopores (pores between 2 and 50 nm). Silica monoliths therefore possess higher surface areas than polymer monoliths and have mainly been utilied for the separation of small molecules, where excellent separation performance has been demonstrated. Polymer monoliths on the other hand have been shown to provide excellent separation performance for larger molecules, where the convective transport of these molecules is unhindered by the presence of mesopores. Polymer and silica monoliths can be prepared in a variety of ways, resulting in varied surface chemistries and flow properties, thus allowing their application in a wide range of chromatographic modes, including reversed‐phase (RP), ion‐exchange, hydrophilic interaction, and hydrophobic interaction chromatography (HIC), to name a few. This has allowed for the separation of a variety of analytes, from small molecules such as alkylbenzenes, to larger molecules such as peptides and proteins. They have also found application for the separation of DNA. In recent years, significant attention has been focused on improving and understanding the behavior of these materials and alternative synthetic routes for their preparation.

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Direct chromatographic capture of enzyme from crude homogenate using immobilized metal affinity chromatography on a continuous supermacroporous adsorbent

Cited by 245 publications

References 11 publications

Detachment of affinity-captured bioparticles by elastic deformation of a macroporous hydrogel

Detachment of affinity-captured bioparticles by elastic deformation of a macroporous hydrogel

Binding mitochondria to cryogel monoliths allows detection of proteins specifically released following permeability transition

Monolithic High‐Performance Liquid Chromatography Columns

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