PREPARATION OF MOLECULARLY IMPRINTED POLYMERS

• US 20120270964A1
• Filed: 12/01/2010
• Published: 10/25/2012
• Est. Priority Date: 12/01/2009
• Status: Active Grant

• Current Assignee: MIP Diagnostics Limited
• Original Assignee: Cranfield University
• Inventors: Piletsky, Sergey Anatollyovich, Guerreiro, Antonio Ricardo Leonardo, Whitcombe, Michael James
• Granted Patent: US 10,189,934 B2
• Time in Patent Office: Days
• US Class Current: 522/181
• CPC Class Codes: B01J 20/268 Polymers created by use of a template, e.g. molecularly imprinted polymers
• CPC Class Codes: C08F 2438/03 Use of a di- or tri-thiocarbonylthio compound, e.g. di- or tri-thioester, di- or tri-thiocarbamate, or a xanthate as chain transfer agent, e.g . Reversible Addition Fragmentation chain Transfer [RAFT] or Macromolecular Design via Interchange of Xanthates [MADIX]
• CPC Class Codes: C08F 293/005 using free radical "living" or "controlled" polymerisation, e.g. using a complexing agent
• CPC Class Codes: G01N 2600/00 Assays involving molecular imprinted polymers/polymers created around a molecular template

Abstract

A polymerization process, preferably a living radical polymerisation process, is carried out in the presence of a template substance, to produce s molecularly imprinted polymers (“MIPs”). It is controlled so that the products are quite small (500-10⁶ Daltons) so that they are soluble or form colloidal suspensions. The template substances are immobilized templates, which can be re-used. Immobilized templates are also useful for purification of MIP solutions/suspensions by affinity chromatography.

Claims

1. A process for the preparation of a molecularly imprinted polymer (“

MIP”

) in the form of a solution or colloidal suspension of MIP particles, comprising the steps of;

(a) providing a carrier substance having a template material immobilized on it so as to be exposed at a surface;

(b) providing a polymerizable composition in contact with said surface;

(c) effecting controlled polymerization of said polymerizable composition in contact with said surface by using a method selected from radical polymerization, optionally controlled living radical polymerization (LRP);

living anionic polymerization;

living cationic polymerization; and

controlled polycondensation;

iniferter-mediated polymerization, nitroxide-mediated polymerization (NMP), atom-transfer radical polymerization (ATRP) and is reversible addition-fragmentation chain-transfer polymerization (RAFT), said polymerization being terminated when MIP particles capable of forming (1) a solution of MIP particles with molecular masses in the range of 500 to 1,000,000 Daltons or (2) a stable colloidal suspension of MIP particles having particle sizes less than 1 micrometer have been formed; and

(d) separating said MIP particles from said surface.

2. A process according to claim 1 wherein said carrier substance is in the form of beads, the surface of waveguides, fibers, optical fibers, membranes, or capillaries.

3. A process according to claim 1 in which said carrier substance is a polymer resin, a polysaccharide, a glass or a metal surface.

4. A process according to claim 1 in which, after step (d) of separating said MIP particles from said surface, said carrier substance with said template material immobilized on it is re-used in steps (b), (c) and (d).

5. A process according to claim 1 where said polymerizable composition contains one or more monomers selected from vinyl monomers, allyl monomers, acetylenes, acrylates, methacrylates, acrylamides, methacrylamides, chloroacrylates, itaconates, trifluoromethylacrylates, derivatives of amino acids, nucleosides, nucleotides, and carbohydrates.

6. A process according to claim 5 wherein said polymerizable composition also contains at least one cross-linker selected from ethylene glycol dimethacrylate, trimethylolpropane trimethacrylate, divinylbenzene, methylene bisacrylamide, ethylene bisacrylamide and N,N′

-bisacryloylpiperazine.

7. A process according to claim 6 wherein, after step (c), poorly bound and low affinity materials are removed from the said immobilized template material by washing, whereafter in step (d), stronger elution conditions are applied for the elution and collection of high affinity particles.

8. A process according to claim 6 wherein in step (c), polymerization is initiated by heating, UV or visible light irradiation, microwave irradiation, electropolymerization, oxidation, or adding catalyst.

10. A process according to claim 9 in which said carrier substance used in step (e) is as defined in claim 2, and is the same as or different from the carrier substance used in step (c).

11. A process according to claim 9 in which said carrier substance used in step (e) is as defined in claim 3, and is the same as or different from the carrier substance used in step (c).

9. A process for the preparation of a molecularly imprinted polymer (“

MIP”

) in the form of a solution or colloidal suspension of MIP particles, comprising is the steps of;

(a) providing a carrier substance having a template material immobilized on it so as to be exposed at a surface;

(b) providing a polymerizable composition in contact with said surface;

(c) effecting controlled polymerization of said polymerizable composition in contact with said surface by using a method selected from radical polymerization, optionally controlled living radical polymerization (LRP);

living anionic polymerization;

living cationic polymerization; and

controlled polycondensation;

iniferter-mediated polymerization, nitroxide-mediated polymerization (NMP), atom-transfer radical polymerization (ATRP) and reversible addition-fragmentation chain-transfer polymerization (RAFT), said polymerization being terminated when MIP particles capable of forming (1) a solution of MIP particles with molecular masses in the range of 500 to 1,000,000 Daltons or (2) a stable colloidal suspension of MIP particles having particle sizes less than 1 micrometer have been formed;

(d) separating said MIP particles from said surface; and

(e) purifying the solution or colloidal suspension of separated MIP particles by contacting it with a carrier substance having said template material immobilized on it so as to be exposed at a surface, so that said MIP particles bind to said immobilized template material, unbound material is separated from said carrier substance, and subsequently MIP particles are recovered from the immobilized template material to form a purified solution or colloidal suspension.

12. A process according to claim 9 where said polymerizable composition contains one or more monomers selected from vinyl monomers, allyl monomers, acetylenes, acrylates, methacrylates, acrylamides, methacrylamides, chloroacrylates, itaconates, trifluoromethylacrylates, derivatives of amino acids, nucleosides, nucleotides, and carbohydrates.

13. A process according to claim 12 wherein said polymerizable composition also contains at least one cross-linker selected from ethylene glycol dimethacrylate, trimethylolpropane trimethacrylate, divinylbenzene, methylene bisacrylamide, ethylene bisacrylamide and N,N′

-bisacryloylpiperazine.

14. A process according to claim 13 wherein said step (d) of separating said MIP particles from said surface is effected by means of heating, changing solution pH, changing ionic strength, or through the addition of urea, guanidine, or a substance which interacts with the template more strongly than does the MIP.

15. A process according to claim 13 wherein said step (c) of effecting controlled polymerization employs sequential polymerization wherein said polymerization composition provided in step (b) is used for a first polymerization step, after which a different polymerisable composition is provided and a further polymerization step is carried out to produce modified MIP particles that are still capable of forming a solution or colloidal suspension.

16. A process according to claim 9 wherein, after step (c), poorly bound and low affinity materials are removed from the said immobilized template material by washing, whereafter in step (d), stronger elution conditions are applied for the elution and collection of high affinity particles.

17. A process according to claim 9 wherein in step (c), polymerization is initiated by heating, UV or visible light irradiation, microwave irradiation, electropolymerization, oxidation, or adding catalyst.

18. A process for the preparation of a molecularly imprinted polymer (“

MIP”

) in the form of a solution or colloidal suspension of MIP particles, comprising the steps of;

(a) providing a carrier substance having a template material immobilized on it so as to be exposed at a surface;

(b) providing a polymerizable composition in contact with said surface, wherein said polymerizable composition contains one or more monomers selected from vinyl monomers, allyl monomers, acetylenes, acrylates, methacrylates, acrylamides, methacrylamides, chloroacrylates, itaconates, trifluoromethylacrylates, derivatives of amino acids, nucleosides, nucleotides, and carbohydrates and also contains at least one cross-linker selected from ethylene glycol dimethacrylate, trimethylolpropane trimethacrylate, divinylbenzene, methylene bisacrylamide, ethylene bisacrylamide and N,N′

-bisacryloylpiperazine;

(c) effecting controlled polymerization of said polymerizable composition in contact with said surface by using a method selected from radical polymerization, optionally controlled living radical polymerization (LRP);

living anionic polymerization;

living cationic polymerization; and

controlled polycondensation;

iniferter-mediated polymerization, nitroxide-mediated polymerization (NMP), atom-transfer radical polymerization (ATRP) and reversible addition-fragmentation chain-transfer polymerization (RAFT), said polymerization being initiated by heating, UV or visible light irradiation, microwave irradiation, electropolymerization, oxidation, or adding catalyst and being terminated when MIP particles capable of forming (1) a solution of MIP particles with molecular masses in the range of 500 to 1,000,000 Daltons or (2) a stable colloidal suspension of MIP particles having particle sizes less than 1 micrometer have been formed; and

(d) separating said MIP particles from said surface by means of heating, changing solution pH, changing ionic strength, or through the addition of urea, guanidine, or a substance which interacts with the template more strongly than does the MIP.

19. A process according to claim 18 wherein said carrier substance is in the form of beads, the surface of waveguides, fibers, optical fibers, membranes, or capillaries.

20. A process according to claim 18 in which said carrier substance is a polymer resin, a polysaccharide, a glass or a metal surface.