Ligand functionalized polymers
Ligand functionalized substrates, methods of making ligand functionalized substrates, and methods of using functionalized substrates are disclosed.
1. A method of separating a target biological species from a fluid, wherein the target biological species is neutral or negatively charged, and wherein the fluid is a cell culture or fermentation broth, comprising:
a. contacting the fluid with a suspension or solution of water-soluble or water-dispersible aminopolymer functionalized with pendent or catenary guanidinyl groups represented by the following general formula:
wherein
R 2 is H, C 1 -C 12 alkyl, C 5 -C 12 (hetero)aryl, or a residue of the polymer chain;
each R 3 is independently H, C 1 -C 12 alkyl, or C 5 -C 12 (hetero)aryl,
each R 4 is H, C 1 -C 12 alkyl or alkylene, C 5 -C 12 (hetero)aryl or (hetero)arylene, cyano, or —C(═NH)—N(R 2 )-Polymer,
n is 1 or 2; and
“Polymer-N(R 2 )—” is the linkage formed between an amine group of the polyamino polymer and guanylating agent;
b. forming a complex in the fluid that comprises the aminopolymer functionalized with guanidinyl groups and the target biological species; and
c. separating the complex from the fluid;
wherein the target biological species is selected from biomacromolecules and microbiological species.
2. The method according to claim 1 , wherein the aminopolymer functionalized with guanidinyl groups is prepared from aminopolymer selected from the group consisting of polyethylenimine, polylysine, polyaminoamides, polyallylamine, polyvinylamine, polydimethylamine-epichlorohydrin-ethylenediamine, polyaminosiloxanes and dendrimers formed form polyamidoamine (PAMAM) and polypropylenimine.
3. The method according to claim 2 , wherein the aminopolymer is polyethylenimine or polyallylamine.
4. The method according to claim 1 , wherein 0.1 to 20 mole percent of the available amino groups of the aminopolymer functionalized with guanidinyl groups are alkylated and/or acylated.
5. The method according to claim 1 , wherein 2 to 10 mole percent of the available amino groups of the aminopolymer functionalized with guanidinyl groups are alkylated and/or acylated.
6. The method according to claim 1 , wherein the aminopolymer functionalized with guanidinyl groups is salt tolerant.
7. The method according to claim 6 , wherein 0.1 to 20 mole percent of the available amino groups of the aminopolymer functionalized with guanidinyl groups are alkylated and acylated.
8. The method of claim 1 , wherein sufficient guanylation agent is used to prepare the aminopolymer functionalized with guanidinyl groups to result in 2.5 to 50 mole percent of the available amino groups of the aminopolymer being functionalized with guanidinyl groups.
9. The method of claim 1 , wherein the target biological species is cells or cell debris.
10. The method according to claim 1 , wherein after separation step (c), the fluid comprises protein or enzyme.
11. The method according to claim 1 , wherein after separation step (c), the fluid comprises antibodies.
12. The method according to claim 1 , wherein the salt concentration of the fluid is between 0 and 300 millimolar.
13. The method according to claim 12 , wherein the salt concentration of fluid is between 150 to 300 millimolar.
14. The method according to claim 1 , wherein the salt concentration of the fluid is at least 150 mM.
15. The method according to claim 12 , wherein the target biological species remaining disposed in the fluid after separation step (c) is less than 50% of its concentration in the fluid in step (a).
16. The method according to claim 12 , wherein the target biological species remaining disposed in the fluid after separation is less than 10% of its concentration in the fluid in step (a).
17. The method according to claim 12 , wherein the target biological species remaining disposed in the fluid after separation is less than 1% of its concentration in the fluid in step (a).
18. The method according to claim 1 wherein the fluid in has a conductivity in the range of 15 to 30 mS/cm.
19. The method according to claim 18 , wherein the target biological species remaining disposed in the fluid after separation step (c) is less than 50% of its concentration in the fluid in step (a).
20. The method according to claim 1 , wherein the amount of aminopolymer functionalized with guanidinyl groups relative to the amount of target biological species is 0.01% to 100% by weight.
21. The method according to claim 20 , wherein the amount of aminopolymer functionalized with guanidinyl groups relative to the amount of target biological species is 0.1%-10% by weight.
22. The method of claim 1 wherein the aminopolymer functionalized with guanidinyl groups is present in amounts of 0.01 to 5000 micrograms/mL of fluid.
23. The method according to claim 1 , wherein the complex precipitates from the fluid.