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231223s2010 xx |||||o 00| ||eng c |
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|a 10.1021/la902135t
|2 doi
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|a pubmed24n0636.xml
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|a (DE-627)NLM190652756
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|a (NLM)19678624
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|a DE-627
|b ger
|c DE-627
|e rakwb
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|a eng
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|a Chung, Wai Keen
|e verfasserin
|4 aut
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|a Utilization of lysozyme charge ladders to examine the effects of protein surface charge distribution on binding affinity in ion exchange systems
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|c 2010
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|a Text
|b txt
|2 rdacontent
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|a ƒaComputermedien
|b c
|2 rdamedia
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|a ƒa Online-Ressource
|b cr
|2 rdacarrier
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|a Date Completed 11.03.2010
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|a Date Revised 21.11.2013
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|a published: Print
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|a Citation Status MEDLINE
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|a A lysozyme library was employed to study the effects of protein surface modification on protein retention and to elucidate preferred protein binding orientations for cation exchange chromatography. Acetic anhydride was used as an acetylating agent to modify protein surface lysine residues. Partial acetylation of lysozyme resulted in the formation of a homologous set of modified proteins with varying charge densities and distribution. The resulting protein charge ladder was separated on a cation exchange column, and eluent fractions were subsequently analyzed using capillary zone electrophoresis and direct infusion electrospray ionization mass spectrometry. The ion exchange separation showed a significant degree of variation in the retention time of the different variants. Several fractions contained coelution of variants, some with differing net charge. In addition, several cases were observed where variants with more positive surface charge eluted from the column prior to variants with less positive charge. Enzymatic digest followed by mass spectrometry was performed to determine the sites of acetylation on the surface of the variants eluting in various fractions. Electrostatic potential maps of these variants were then generated to provide further insight into the elution order of the variants
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|a Journal Article
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|a Research Support, U.S. Gov't, Non-P.H.S.
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|a Acetic Anhydrides
|2 NLM
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|a acetic anhydride
|2 NLM
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|a 2E48G1QI9Q
|2 NLM
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|a Muramidase
|2 NLM
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|a EC 3.2.1.17
|2 NLM
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|a Lysine
|2 NLM
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|a K3Z4F929H6
|2 NLM
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1 |
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|a Evans, Steven T
|e verfasserin
|4 aut
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1 |
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|a Freed, Alexander S
|e verfasserin
|4 aut
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1 |
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|a Keba, James J
|e verfasserin
|4 aut
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|a Baer, Zachary C
|e verfasserin
|4 aut
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|a Rege, Kaushal
|e verfasserin
|4 aut
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| 700 |
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|a Cramer, Steven M
|e verfasserin
|4 aut
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| 773 |
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|i Enthalten in
|t Langmuir : the ACS journal of surfaces and colloids
|d 1992
|g 26(2010), 2 vom: 19. Jan., Seite 759-68
|w (DE-627)NLM098181009
|x 1520-5827
|7 nnns
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| 773 |
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|g volume:26
|g year:2010
|g number:2
|g day:19
|g month:01
|g pages:759-68
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|u http://dx.doi.org/10.1021/la902135t
|3 Volltext
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