Graft copolymer functionalized article
View Patent ↗Guanidinyl ligand-functionalized polymers, methods of making the same, and substrates bearing a grafted coating of the ligand-functional polymers are described. The grafted polymer has the requisite affinity for binding neutral or negatively charged biomaterials, such as cells, cell debris, bacteria, spores, viruses, nucleic acids, endotoxins and proteins, at pH's near or below the pI's of the biomaterials.
1. A method of preparing a ligand functional substrate comprising the steps of:
a) providing a substrate,
b) free-radically grafting the substrate in the presence of a Type II photoinitiator of the general formula:
Ar—CO—R 13
in which Ar is a substituted or unsubstituted aryl group having 6 to 12 carbon atoms optionally substituted with a C 1 to C 12 alkyl group, a C 1 to C 12 alkoxy group, or a phenyl group; and
R 13 is a C 1 to C 6 alkyl group, a cycloalkyl group having 3 to 14 carbon atoms, or Ar;
with a guanidinyl-functional (meth)acryloyl monomer of the formula:
wherein
R 1 is H or C 1 -C 4 alkyl;
R 2 is a (hetero)hydrocarbyl group;
each R 3 is independently H or hydrocarbyl;
R 4 is H, C 1 -C 12 alkyl or —N(R 3 ) 2 ;
R 5 is H or hydrocarbyl;
X 1 is —O— or —NR 3 —;
o is 0 or 1, and
n is 1 or 2; and
wherein the substrate is provided with a primer layer disposed on the substrate comprising the reaction product of:
1) a polyamine polymer; and
2) a polyfunctional crosslinking agent for the polyamine polymer.
2. The method of claim 1 wherein the substrate is provided with a primer layer disposed on the substrate comprising the reaction product of:
1) a polyamine polymer;
2) a polyfunctional crosslinking agent for the polyamine polymer; and
3) an amine-reactive monomer having an amine-reactive functional group and an ethylenically unsaturated, polymerizable group, to provide a substrate having a crosslinked polyamine primer layer on the substrate, the primer layer having pendent alkenyl groups.
3. The method of claim 1 further comprising grafting the substrate with a hydrophilic monomer.
4. The method of claim 1 further comprising grafting with a multifunctional (meth)acryloyl monomer.
5. The method of claim 1 wherein the Type II photoinitiator is a benzophenone photoinitiator.
6. The method of claim 1 comprising the step of free-radically grafting the substrate with a monomer mixture comprising:
a) up to 100 parts by weight of guanidinyl monomers,
b) 0 to 90 parts by weight of hydrophilic monomer units, and
c) 0 to about 5 by weight parts by weight of multifunctional (meth)acryloyl monomer wherein the total monomer is 100 parts by weight.
7. The method of claim 6 wherein the monomer mixture comprises about 0.1 wt % to about 60 wt % of monomers in an organic or aqueous solvent.
8. The method of claim 1 wherein the grafted monomers form a grafted copolymer of the formula:
-(M Lig ) y -(M Hydrophil ) x -(M crosslink ) z -, where
(M Hydrophil ) x are hydrophilic monomer units having “x” polymerized monomer units,
(M Lig ) y are ligand functional monomer units having “y” polymerized monomer units,
(M crosslink ) are multifunctional (meth)acrylate monomer units having “z” polymerized monomer units.
9. The method of claim 3 wherein the hydrophilic monomer units comprise poly(oxyalkylene) (meth)acrylate monomer units.
10. The method of claim 9 , wherein the poly(oxyalkylene) (meth)acrylate monomer units are of the formula: CH 2 ═CR 1 —C(O)—X 1 —(CH(R 1 )—CH 2 —O) n —R 1 , wherein each R 1 is independently H or C 1 -C 4 alkyl, X 1 is —O— or —NR 3 —, where R 3 is H or C 1 -C 4 alkyl and n is 2 to 100.
11. The method of claim 1 wherein the substrate comprises a primer layer disposed on the substrate comprising a crosslinked polyamine polymer having ethylenically unsaturated polymerizable groups.
12. The method of claim 11 wherein the crosslinked polyamine polymer having ethylenically unsaturated polymerizable groups is the reaction product of:
1) a polyamine polymer;
2) a polyfunctional crosslinking agent for the polyamine polymer; and
3) a monomer having an amine-reactive functional group and an ethylenically unsaturated polymerizable group.
13. The method of claim 2 wherein the polyamine polymer is selected from the group consisting of polyethylenimine, polylysine, polyaminoamides, polydimethylamine-epichlorohydrin-ethylenediamine, polyaminosiloxanes and dendrimers formed from polyamidoamine (PAMAM) and polypropylenimine.
14. The method of claim 2 wherein said primer layer comprises 0.1 to 5 wt. % of the substrate.
15. The method of claim 8 wherein said (co)polymer comprises 5 to 30 wt. % of the ligand functional substrate.
16. A method of separating a target biological species from a fluid comprising contacting the fluid with the ligand-functionalized substrate prepared by the method of claim 1 whereby a complex comprising the functionalized substrate and the target biological species is formed, and separating the complex; wherein said target biological species is selected from biomacromolecules and microbiological species.
17. The method of claim 16 wherein said biomacromolecules are selected from proteins, enzymes, nucleic acids, and endotoxins.
18. The method of claim 16 wherein said microbiological species is selected from bacteria, viruses, cells, cell debris, and spores.
19. The method of claim 18 wherein the cells are selected from archaea, bacteria, and eucaryota.
20. The method of claim 16 wherein the biological fluid is derived from a cell culture or fermentation process.
21. The method of claim 16 wherein the biological fluid comprises a solution of a purified protein or enzyme after separating the target species.
22. The method of claim 21 wherein the separated target species comprises a purified protein or enzyme.
23. The method of claim 16 wherein the fluid has a salt content of at least 50 millimolar.