On-line concentration of peptides and proteins with the hyphenation of polymer monolithic immobilized metal affinity chromatography and capillary electrophoresis.
An iminodiacetic acid (IDA)-type adsorbent is prepared at the one end of a capillary by covalently bonding IDA to the monolithic rods of macroporous poly(glycidyl methacrylate-co-ethylene dimethacrylate). Cu(II) is later introduced to the support via the interaction with IDA. By this means, polymer monolithic immobilized metal affinity chromatography (IMAC) materials are prepared. With such a column, IMAC for on-line concentration and capillary electrophoresis (CE) for the subsequent analysis are hyphenated for the analysis of peptides and proteins. The reproducibility of such a column has been proved good with relative standard deviations (RSDs) of dead time of less than 5% for injection-to-injection and 12% for column-to-column (n = 3). Through application on the analysis of standard peptides and real protein samples, such a technique has shown promising in proteome study.
Zhang
Lingyi
L
National Chromatographic Research & Analysis Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, PR China.
Zhang
Lihua
L
Zhang
Weibing
W
Zhang
Yukui
Y
D002846:Chromatography, Affinity / Q000379:methods*; D003300:Copper; D019075:Electrophoresis, Capillary / Q000379:methods*; D007098:Imino Acids; D008670:Metals; D008768:Methylmethacrylates; D009862:Online Systems; D010455:Peptides / Q000032:analysis*; D011108:Polymers; D016062:Porosity; D011506:Proteins / Q000032:analysis*; D040901:Proteomics / Q000379:methods
D007098:Imino Acids; D008670:Metals; D008768:Methylmethacrylates; D010455:Peptides; D011108:Polymers; D011506:Proteins; C020347:GMA-EDMA; D003300:Copper; C008109:iminodiacetic acid