Repeatedly usable immobilized pH gradient in a monolithic capillary column.
Glycidyl methacrylate (GMA) and ethylene dimethacrylate (EDMA) were used to synthesize a monolithic capillary column containing reactive epoxy groups. Glutaraldehyde was introduced and linked to the monolith after a process of amination. An aqueous solution of commercial carrier ampholytes (CAs, Ampholine) was focused in such a polymer column. The primary amino groups of CAs reacted with glutaraldehyde along the capillary. CAs were immobilized at different positions in the column according to their isoelectric points (pI), resulting in a monolithic immobilized pH gradient (M-IPG). Isoelectric focusing (IEF) was performed without CAs in such an M-IPG column. Due to the covalent attachment of the CAs this M-IPG can be repeatedly used after its preparation. Good stability, linearity, and reproducibility were obtained.
Yang
Chun
C
Department of Biotechnology, National Chromatographic R. & A. Center, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, China.
Zhu
Guijie
G
Zhang
Lihua
L
Zhang
Weibing
W
Zhang
Yukui
Y
D000586:Amination; D000665:Ampholyte Mixtures / Q000737:chemistry*; D002196:Capillaries / Q000737:chemistry*; D004852:Epoxy Compounds / Q000737:chemistry*; D004853:Epoxy Resins / Q000737:chemistry; D007525:Isoelectric Focusing; D007526:Isoelectric Point; D008689:Methacrylates / Q000737:chemistry*; D011073:Polyamines; D011108:Polymers / Q000737:chemistry*; D018892:Proton-Motive Force
D000665:Ampholyte Mixtures; D004852:Epoxy Compounds; D004853:Epoxy Resins; D008689:Methacrylates; D011073:Polyamines; D011108:Polymers; C002023:Ampholine; C004919:ethylene dimethacrylate; C007870:glycidyl methacrylate