DE3126551A1 - Process for the preparation of materials for the immobilisation of proteins and carbohydrate groups - Google Patents

Abstract

The invention has applications in the area of solid phase techniques and affinity chromatography. The aim of the invention, to provide carrier materials with improved protein-binding properties but also the possibility of binding carbohydrate groups, is achieved by functionalising the surfaces of hydrophobic materials by amphiphilic substances (e.g. phospho-, sphingo- and/or glycosphingolipids) using essentially non-polar solvents in such a way that, after removal of the solvent, the hydrophilic portion of the amphiphile, which is bonded to the material by van der Waal's forces, is available for further reactions of a covalent and/or adsorptive nature with proteins and/or carbohydrate groups in aqueous solution.

Description

3126

Rolf Siegel - Z. * Tannenweg 3

6507 Ingelheim

Manufacturing process for materials for immobilizing proteins and carbohydrate groups This invention relates to a manufacturing method for materials for immobilizing proteins and carbohydrate groups and is mainly used where protein and / or carbohydrate molecules are on a solid Base are fixed, so in the field of "solid phase" techniques, such as the matrix-bound enzymes ("matrix bound enzymes", "immobilized enzymes") and affinity chromatography.

Ks is known to be due to polystyrene and polypropylene Ionic and hydrophobic interactions adsorb protein molecules. The use of this is based on this fact Plastics as a solid base ("solid phase"), for example as "coated tubes" coated with antibodies Radioimmuno3ssays or as coated with antigens! ■■ ikrotiterplstten with 6PIT ("solid phase immune test").

The protein binding capacity of these plastics is low: Pesce, AJ, JJ. J. Ford, K. Gaizutis and VE Pollack: iochem. Eiophys. acta 492, 399 (1977), as well as Ohristensen, P., op. Johansson and V. Kielsen: J. Iramunol. Kethods 23, 23 (197S). - «From the work of öslonen and Vaheri (rfalonen, E.-K. and A. Vaheri: J. Immunol, 1.eühocs 30,? 0Q (1479)) it furthermore states that: · the VorDehonalur :. -of tolystyrene with hydrochloric acid, / · ethanol, Ätivmol / Vtceton- • iemifichen and Cilutar -'- 'idehyd koine improvement of the Proüi.xü i nd u η ¹ V s k a ρ a ζ it ^: ite rg ib t.

An improvement in the protein loading capacity, above all the binding of carbohydrate groups to the surfaces hydrophobic Ι'-materials, such as (cross-linked) polystyrene, According to the invention, polypropylene or nylon is achieved by targeting functional parts on the corresponding surfaces Groups such as Phosphate, primary amine, carboxyl,

(3arüa * id, thiol, auarternary ammonium groups, etc., above also carbohydrate groups are applied in that

BATH ORIGINAL

Compounds, the molecules of which have both a strongly hydrophobic and a (highly) polar component, the latter in turn being the carrier of the respective functional group, preferably compounds from the class of phospholipoids, apinrcolioids or glycosphingolipoids, as non-substances or as mixtures, essentially in one non-polar solvent or can be dissolved or suspended Lösungsmittelgernisch, and that this solution or _"A .Suspension is brought into intimate contact with the surfaces of the, respective hydrophobic material, and that subsequently the solvent (mixture), preferably by evaporation at reduced pressure, again is completely removed. With this procedure, the compounds mentioned - ideally with the formation of a monomolecular layer - with their hydrophobic components are firmly attached to the surfaces of the hydrophobic material by means of van der Vaals' forces. The (highly) polar components of the compounds, which in turn are carriers of the respective functional groups, are now available on the surface of the material for further reactions of a covalent or adsorptive type with proteins or carbohydrate groups are hydrophobic starting materials as compared to other solid support used f-iaterialien such as dextran, or cellulose kgarose a) tiicht degraded by microorganisms and b) low cost in a variety UNTT rGchiüolic'ier-i-i3terial ^1; _t y!; s are available, zi crosslinked / non-crosslinked polystyrene with different pore sizes, inechanical special cha''ten and standards ( ^; 'be: icis ^{: i} ocier "latte surfaces).' - The main advantage, however, lies in this that a large number of different functional groups can be applied to the surfaces of the hydrophonic materials in one operation. These materials can be designed, for example by applying a cell membrane extract, in such a way that the surfaces largely correspond to the natural model. Immobilization of enzymes on such surfaces then corresponds far more to physiological conditions than the binding via only one functional group, as has happened so far with the known materials for "solid phase" techniques.

BATH ORIGINAL

Claims (1)

Rolf Siegel Tannenweg 3 6507 Ingelheim Claim Manufacturing process for materials for immobilizing proteins and carbohydrate groups, characterized in that that the surfaces of hydrophobic materials such as (crosslinked) polystyrene, polypropylene or nylon are functional Groups such as phosphate, primary aniine, carboxyl, Carbamide, thiol, quaternary ammonium groups, etc., but Carbohydrate groups obtained in that compounds whose molecules have both a strongly hydrophobic and a have a (highly) polar fraction, the latter in turn being a carrier of the respective functional group, preferably Compounds from the class of phospholipids, sphingolipids or glycosphingolipids, as pure substances or as mixtures, dissolved or suspended in an essentially non-polar solvent or solvent mixture be, and that this solution or suspension with the surfaces of the respective hydrophobic material in intimate Is brought into contact, and that then the solvent mixture), preferably by evaporating at reduced jJruck, is completely removed again. ¹ jsis -cscnai'fene such material is now available for further ■■ '.eckt ion kovlenter or adsorptive -irt with proteins or i.onlenhydrat: _-: rui: oen for disposal. BATHROOM OHlGINAL