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231224s2012 xx |||||o 00| ||eng c |
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|a 10.1021/la300481e
|2 doi
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|a eng
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|a Bhattacharjee, Somnath
|e verfasserin
|4 aut
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|a Formation of high-capacity protein-adsorbing membranes through simple adsorption of poly(acrylic acid)-containing films at low pH
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|c 2012
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|a Text
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|a ƒaComputermedien
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|2 rdamedia
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|a ƒa Online-Ressource
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|a Date Completed 17.08.2012
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|a Date Revised 15.03.2024
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|a published: Print-Electronic
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|a Citation Status MEDLINE
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|a Layer-by-layer polyelectrolyte adsorption is a simple, convenient method for introducing ion-exchange sites in porous membranes. This study demonstrates that adsorption of poly(acrylic acid) (PAA)-containing films at pH 3 rather than pH 5 increases the protein-binding capacity of such polyelectrolyte-modified membranes 3-6-fold. The low adsorption pH generates a high density of -COOH groups that function as either ion-exchange sites or points for covalent immobilization of metal-ion complexes that selectively bind tagged proteins. When functionalized with nitrilotriacetate (NTA)-Ni(2+) complexes, membranes containing PAA/polyethylenimine (PEI)/PAA films bind 93 mg of histidine(6)-tagged (His-tagged) ubiquitin per cm(3) of membrane. Additionally these membranes isolate His-tagged COP9 signalosome complex subunit 8 from cell extracts and show >90% recovery of His-tagged ubiquitin. Although modification with polyelectrolyte films occurs by simply passing polyelectrolyte solutions through the membrane for as little as 5 min, with low-pH deposition the protein binding capacities of such membranes are as high as for membranes modified with polymer brushes and 2-3-fold higher than for commercially available immobilized metal affinity chromatography (IMAC) resins. Moreover, the buffer permeabilities of polyelectrolyte-modified membranes that bind His-tagged protein are ~30% of the corresponding permeabilities of unmodified membranes, so protein capture can occur rapidly with low-pressure drops. Even at a solution linear velocity of 570 cm/h, membranes modified with PAA/PEI/PAA exhibit a lysozyme dynamic binding capacity (capacity at 10% breakthrough) of ~40 mg/cm(3). Preliminary studies suggest that these membranes are stable under depyrogenation conditions (1 M NaOH)
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|a Journal Article
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|a Research Support, N.I.H., Extramural
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|a Acrylic Resins
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|a Membranes, Artificial
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|a Nylons
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|a Organometallic Compounds
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|a Polyamines
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|a Proteins
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|a polyallylamine
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|a 30551-89-4
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|a nickel nitrilotriacetic acid
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|a Nitrilotriacetic Acid
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|a KA90006V9D
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|a Dong, Jinlan
|e verfasserin
|4 aut
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|a Ma, Yiding
|e verfasserin
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|a Hovde, Stacy
|e verfasserin
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|a Geiger, James H
|e verfasserin
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|a Baker, Gregory L
|e verfasserin
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|a Bruening, Merlin L
|e verfasserin
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|i Enthalten in
|t Langmuir : the ACS journal of surfaces and colloids
|d 1992
|g 28(2012), 17 vom: 01. Mai, Seite 6885-92
|w (DE-627)NLM098181009
|x 1520-5827
|7 nnns
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|g volume:28
|g year:2012
|g number:17
|g day:01
|g month:05
|g pages:6885-92
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|u http://dx.doi.org/10.1021/la300481e
|3 Volltext
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