Peptide recognition via hierarchical imprinting

Silica particles containing immobilised peptidic templates have been used for the generation of hierarchically imprinted polymers. The pores of the silica mould were filled with a mixture of monomers/initiator and polymerised, followed by dissolution of the silica template. This method leaves behind imprinted polymers with binding sites located at the surface, which are capable of recognising larger molecules with the same immobilised epitope. All the products resulting from solid-phase synthesis of peptides were characterised by elemental analysis, FT-IR spectroscopy and fluorescence microscopy. The hierarchically imprinted polymers generated from these products were characterised by elemental analysis, FT-IR spectroscopy, fluorescence microscopy, scanning electron microscopy (SEM) and nitrogen adsorption, providing evidence concerning the reproducibility of each step. The chromatographic properties of the materials have been investigated and the advantages of the immobilisation method have been proven. The materials exhibit selectivity for their templates and other structurally related dipeptides. Furthermore, the polymers proved to be capable of recognising larger peptides containing the immobilised sequence.

[1]  Börje Sellergren,et al.  Molecularly imprinted polymers : man-made mimics of antibodies and their applications in analytical chemistry , 2001 .

[2]  E. Carlier,et al.  Functional polymers supported on porous silica. II.Radical polymerization of vinylbenzyl chloride from grafted precursors , 1992 .

[3]  Maria-Magdalena Titirici,et al.  Hierarchically Imprinted Stationary Phases: Mesoporous Polymer Beads Containing Surface-Confined Binding Sites for Adenine , 2002 .

[4]  R. B. Merrifield,et al.  Selective deprotection of the N.alpha.-tert-butyloxycarbonyl group in solid phase peptide synthesis with chlorotrimethylsilane in phenol , 1993 .

[5]  S. Pennycook,et al.  Hierarchically Imprinted Sorbents for the Separation of Metal Ions , 2000 .

[6]  B. Sellergren,et al.  Influence of polymer morphology on the ability of imprinted network polymers to resolve enantiomers , 1993 .

[7]  E. Vulfson,et al.  Surface imprinting of cholesterol on Submicrometer core-shell emulsion particles , 2001 .

[8]  G. Wulff,et al.  Enzyme-like catalysis by molecularly imprinted polymers. , 2002, Chemical reviews.

[9]  N. Minoura,et al.  Molecular imprinting: synthesis of polymer particles with antibody-like binding characteristics for glucose oxidase. , 1996, Biochemical and biophysical research communications.

[10]  A. Martín‐Esteban,et al.  Highly selective fenuron-imprinted polymer with a homogeneous binding site distribution prepared by precipitation polymerisation and its application to the clean-up of fenuron in plant samples , 2003 .

[11]  P. Voort,et al.  Characterization and Chemical Modification of the Silica Surface , 1995 .

[12]  Toshifumi Takeuchi,et al.  Solid-phase Extraction of a Triazine Herbicide Using a Molecularly Imprinted Synthetic Receptor , 1997 .

[13]  Stellan Hjertén,et al.  Gels mimicking antibodies in their selective recognition of proteins , 1997 .

[14]  K. Mosbach,et al.  The Use of Immobilized Templates-A New Approach in Molecular Imprinting. , 2000, Angewandte Chemie.

[15]  Maria-Magdalena Titirici,et al.  Hierarchical Imprinting Using Crude Solid Phase Peptide Synthesis Products as Templates , 2003 .