Synthesis and application of novel phenylboronate affinity materials based on organic polymer particles for selective trapping of glycoproteins

Journal of Separation Science - Tập 32 Số 10 - Trang 1673-1685 - 2009
Beatrix Preinerstorfer1, Michael Lämmerhofer1, Wolfgang Lindner1
1Christian Doppler Laboratory for Molecular Recognition Materials, Department of Analytical Chemistry and Food Chemistry, University of Vienna, Vienna, Austria. Fax: 43‐1‐4277‐9523

Tóm tắt

AbstractWe report on synthesis concepts for the fabrication of various novel phenylboronate affinity materials based on polymethacrylate epoxy beads (Fractogel® EMD Epoxy (M) 40–90 μm) and the testing of these functionalized polymer particles for selective trapping of a glycoprotein from a standard mixture containing a glycosylated and a nonglycosylated protein. Two inherently different approaches for the functionalization of the bare beads with boronate groups have been elucidated. In the first, the epoxy residues of the polymer particles were converted into reactive thiol groups which were subsequently used as anchor moieties for the immobilization of 4‐vinylphenylboronic acid by radical addition or radical polymerization reaction. Three different ways for the generation of sulfhydryl groups have been examined leading to materials with distinct linker chemistries. In the second and more straightforward approach, the epoxy groups were reacted with 4‐mercaptophenylboronic acid. The novel materials were thoroughly characterized by (i) quantitation of the sulfur content by elemental analysis, (ii) reactive sulfhydryls were determined in a photospectrometric assay, (iii) boron content was measured by inductively coupled plasma‐atomic emission spectrometry, and (iv) the amount of reactive boronate groups was evaluated in a fast binding assay employing adenosine as test compound. A maximum concentration of 1.2 mmol boronate groups per gram dry beads could be achieved by the presented synthesis routes. Employing the novel phenylboronate affinity materials in capture and release experiments in the batch mode, a standard glycoprotein, viz. transferrin (Tf) from human serum was separated from a nonglycosylated protein, BSA. A commercial boronate affinity material based on 3‐aminophenylboronic acid modified agarose gel was employed as reference material and was found to perform significantly worse compared to the herein presented novel polymethacrylate particles.

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Tài liệu tham khảo

10.1016/S0040-4020(02)00489-1

10.1021/ac0709089

10.1016/0378-4347(96)00006-0

10.1016/S0378-4347(01)00196-7

10.1016/S1570-0232(02)00162-9

10.1016/S0168-1656(02)00357-7

10.1007/s00216-007-1627-y

10.1365/s10337-008-0677-4

10.1074/jbc.273.47.31125

10.1021/pr070112q

10.1002/jhrc.1240150716

10.1093/oxfordjournals.jbchem.a022243

10.1016/0021-9673(86)80009-7

10.1002/(SICI)1097-0290(19971105)56:3<345::AID-BIT13>3.0.CO;2-F

10.1023/A:1009223615607

10.1002/jssc.200500257

10.1021/ac702057u

10.1021/jo020745i

10.1186/1471-2091-8-25

10.1016/j.bmcl.2003.10.017

10.1016/j.ab.2007.09.001

10.1002/elps.200800178

Kaiser C. Segui‐Lines G. D'Amaral J. C. Ptolemy A. S. Britz‐McKibbin P. Chem. Commun.2008 338–340.

10.1039/b609051f

10.1016/j.chroma.2003.10.023

10.1021/ja026658p

10.1021/ja027919y

10.1016/j.snb.2005.07.011

10.1016/j.jelechem.2006.04.021

10.1016/S0960-894X(02)00767-9

10.1002/elan.200302809

10.1002/chem.200600442

10.1021/ja073939u

10.1016/S1381-5148(01)00056-6

10.1016/j.ab.2004.05.051

10.1016/j.colsurfa.2003.11.007

10.1002/chem.200600342

10.1016/S0956-5663(01)00187-7

10.1002/rcm.2793

10.1021/la050171n

10.1021/la7007733

10.1021/ac702588b

10.1021/pr0702767

10.1002/adfm.200800215

10.1016/j.chroma.2005.03.038

10.1016/j.aca.2004.11.017

10.1016/0021-9673(92)85535-2

10.1016/S0021-9673(00)01106-7

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Thông tin xuất bản

Journal of Separation Science

Tập 32 Số 10

1673-1685

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