Selective Sorbents - Silica-immobilized affinity ligands and molecularly imprinted polymers

Research output: ThesisDoctoral Thesis (compilation)

Abstract

The development of selective sorbents for the separation of various molecules, preferentially biomolecules, is described in this work. Affinity ligands were immobilized on small silica particles and used in high-performance liquid affinity chromatography (HPLAC). With this technique, it is possible to perform fast, selective and efficient bioseparations. Methods were developed for the preparation of silica-immobilized phenyl boronic acid and its use in general ligand affinity chromatography of, e.g., carbohydrates, catechols and nucleotides. Cibacron Blue is another general ligand, with selectivity for many biomolecules, such as enzymes and binding proteins. It was used for the HPLAC separation of proteins such as kinases from yeast extracts. Automatic detection was performed using an on-line enzyme assay system.

Molecular imprinting was employed for the preparation of various selective sorbents. To improve the performance, a composite technology was developed in which small rigid particles of silica or TRIM (trimethylolpropane trimethacrylate) were employed for the anchoring of molecularly imprinted polymers. Proteins were for the first time used as templates in molecular imprinting. The boronate interaction with sialic acid moieties of the glycoprotein transferrin was employed for molecular imprinting in aqueous environments using polymerizable organic silanes on the surface of silica. Bovine pancreatic ribonuclease A, with exposed surface histidines was imprinted using monomers capable of forming metal chelates with the protein. Finally, the advantages of the various technologies described in this work will be discussed, as well as current trends and future prospects.

Details

Authors
  • Magnus Glad
Organisations
Research areas and keywords

Subject classification (UKÄ) – MANDATORY

  • Biochemistry and Molecular Biology

Keywords

  • Molecular imprinting, HPLAC, composites, TRIM, Biokemi, Biochemistry, Metabolism, affinity chromatography
Original languageEnglish
QualificationDoctor
Awarding Institution
Supervisors/Assistant supervisor
  • [unknown], [unknown], Supervisor, External person
Award date2002 Oct 11
Publisher
  • Magnus Glad, Nyckelkroken 64, SE-226 47 Lund, Sweden,
Print ISBNs91-628-5396-1
Publication statusPublished - 2002
Publication categoryResearch

Bibliographic note

Defence details Date: 2002-10-11 Time: 10:15 Place: Kemicentrum Sal B, 10.15 External reviewer(s) Name: Janson, Jan-Christer Title: Prof. Affiliation: Uppsala University, Uppsala --- Article: I High-Performance Liquid Affinity Chromatography of Nucleosides, Nucleotides and Carbohydrates with Boronic Acid-Substituted Microparticulate Silica, Magnus Glad, Sten Ohlson, Lennart Hansson, Mats-Olle Månsson and Klaus Mosbach, Journal of Chromatography, 200 (1980) 254-260 Article: II Boronic Acid-Silica: A New Tool for the Purification of Catecholic Compounds On-Line with Reversed-Phase High-Performance Liquid Chromatography, Lennart Hansson, Magnus Glad and Christer Hansson, Journal of Chromatography, 265 (1983) 37-44 Article: III High-Performance Liquid Affinity Chromatography of Proteins on Cibacron Blue F3G-A Bonded Silica, Christopher R. Lowe, Magnus Glad, Per-Olof Larsson, Sten Ohlson, David A.P. Small, Tony Atkinson and Klaus Mosbach, Journal of Chromatography, 215 (1981) 303-316 Article: IV Acrylic Polymer Preparations Containing Recognition Sites Obtained by Imprinting with Substrates, Olof Norrlöw, Magnus Glad and Klaus Mosbach, Journal of Chromatography, 299 (1984) 29-41 Article: V Molecularly Imprinted Composite Polymers Based on Trimethylolpropane Trimethacrylate (TRIM) Particles for Efficient Enantiomeric Separations, Magnus Glad, Per Reinholdsson and Klaus Mosbach, Reactive Polymers, 25 (1995) 47-54 Article: VI Use of Silane Monomers for Molecular Imprinting and Enzyme Entrapment in Polysiloxane-Coated Porous Silica. Magnus Glad, Olof Norrlöw, Börje Sellergren, Nils Siegbahn and Klaus Mosbach, Journal of Chromatography, 347 (1985) 11-23 Article: VII An Approach Towards Surface Imprinting Using the Enzyme Ribonuclease A, Maria Kempe, Magnus Glad and Klaus Mosbach, Journal of Molecular Recognition, 8 (1995) 35-39