Microparticles with enhanced covalent binding capacity and their uses
View Patent ↗Disclosed are proteins which are covalently bound to a solid support at a first temperature where they have a first configuration, and then biomolecules are attached to the bound proteins at a higher temperature at which the proteins have more exposed functional groups, each such group being capable of covalently bonding to a biomolecule. The biomolecule can be, for example, a nucleic acid, including an amine functionalized oligonucleotide. The proteins can include, BSA (Bovine Serum Albumin) which can be bound using a reaction with the surface of a tosyl-activated microparticle.
1. A product comprising:
(i) a microparticle;
(ii) a single layer of bovine serum albumin molecules covalently attached to the microparticle;
(iii) oligonucleotides covalently attached to the single layer wherein attachment of the single layer to the microparticle is performed at a pre-selected temperature such that the bovine serum albumin molecules in said single layer attached at said pre-selected temperature display a greater number of available sites for covalent attachment of oligonucleotides than if the single layer was attached to the microparticle at a different temperature.
2. The product of claim 1 wherein the pre-selected temperature is about 65° C.
3. The product of claim 1 wherein the oligonucleotides are bound to the protein molecules through an amide linkage, formed by an 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide reaction.
4. The product of claim 1 wherein the surface of the microparticle is activated with tosyl functional group.
5. The product of claim 1 wherein the oligonucleotide is DNA, RNA, a peptide nucleic acid, a locked nucleic acid, or mixtures thereof.
6. The product of claim 1 wherein the oligonucleotide is modified at its 3′ or 5′ end with a functional group.
7. The product of claim 6 wherein the functional group is a primary amino group.