Magnetic particles as supports for organic synthesis

• US 4,638,032 A
• Filed: 08/10/1984
• Issued: 01/20/1987
• Est. Priority Date: 12/12/1983
• Status: Expired due to Fees

• Current Assignee: Genetics Institute LLC (Pfizer Inc.)
• Original Assignee: Genetics Institute LLC (Pfizer Inc.)
• Inventors: Benner, Steven A.
• Primary Examiner(s): Foelak, Morton
• Assistant Examiner(s): Nutter, Nathan M.
• Application Number:
  US06/640,027
• Time in Patent Office: 893 Days
• Field of Search: 536/27, 536/28, 536/29, 536/18.5, 536/124, 524/785, 252/62.54, 252/62.51, 525/54.1, 525/54.4
• US Class Current: 525/54.11
• CPC Class Codes: B03C 1/01 by addition of magnetic adjuvants
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C07K 1/042 characterised by the nature of the carrier
• CPC Class Codes: Y02P 20/582 Recycling of unreacted starting or intermediate materials

Abstract

A support system for organic synthesis comprising magnetic particles in a dispersion medium covalently attached to functional groups having affinity for polymer subunits, and methods for making and using the support system, e.g. for synthesis of oligodeoxynucleotides and polypeptides.

Claims

1. Oligomer synthesis support comprising a magnetic particle covalently bound to oligomer subunits, said oligomer subunits comprising subunits of deoxyribonucleic acid or ribonucleic acid.

2. The composition of claim 1, wherein said oligomer subunits comprise subunits of deoxyribonucleic acid.

3. The composition of claim 1, wherein said magnetic particles comprise magnetite.

4. The composition of claim 1, wherein said oligomer synthesis support exists as a colloidal suspension in a dispersion medium.

5. The composition of claim 1, wherein said magnetic particles are single domain.

6. The composition of claim 1, wherein the size of said magnetic particles is in the range of 10-100,000 Angstroms.

18. The composition of claim 17 or 1, wherein said linking group is polymethylene.

19. The composition of claim 17 or 1, wherein said linking group is N-trimethylene-carboxamide-dimethylene.

20. The composition of claim 17 or 1, wherein said member Z is a deoxyribonucleic acid, ribonucleic acid or polypeptide subunit.

7. An organic synthesis support that comprises a magnetic particle having reactive sites covalently coupled to functional groups, the remaining reactive sites being blocked against reaction by silyl blocking groups.

8. The composition of claim 7, wherein said magnetic particles comprise magnetite.

9. The composition of claim 7, wherein said magnetic particles are non-porous.

10. The composition of claim 7, wherein said magnetic particles exist as a colloidal suspension in a dispersion medium.

11. The composition of claim 7, wherein said magnetic particles are single domain.

12. The composition of claim 7, wherein said magnetic particles are in the range of 10-100,000 Angstroms.

13. The composition of claim 7, wherein said magnetic particles are in the range of 10-100,000 Angstroms.

14. The composition of claim 7, wherein said functional groups are amino, carboxyl, or hydroxyl.

15. The composition of claim 7, wherein the reactive sites are blocked by reaction with a compound having the following structure:

space="preserve" listing-type="equation">Si(X).sub.n (R).sub.4-n

wherein Si is the silicon of an organosilane;

X is a leaving group and member of the group alkoxy, halogen or amine;

n is an integer having a value of 1 or 2;

R is a member of a group of alkyl, alkenyl, alkynyl or aryl where the group contains at least one carbon atom and does not contain any reactive functionality.

16. The composition of claim 7, wherein said silyl blocking agent is trimethysilylchloride.

17. A cross-linked oligomer synthesis support having the following structure:

space="preserve" listing-type="equation">M--O--Si(X).sub.n [R.sub.m (Y).sub.p ].sub.3-n --Z

wherein M is the surface of a magnetic support particle;

O is the oxygen of the metal oxide or hydroxide;

Si is the silicon of the organosilane linking or coupling agent;

X is a member of the group of alkenyloxy, alkaryloxy, alkynyloxy, alkoxy and halo;

n is an integer with a value of 0, 1 or 2;

is a linking group and member of a group of alkyl, alkenyl, alkynyl or aryl where the group contains more than one carbon atom;

n is an integer with a value of 1, 2 or 3;

Y is a member of the group of amino, hydroxyl or carboxyl which will covalently combine with an oligomer subunit or precursor;

p is an integer with a value of at least 1;

Z is an initial oligomer subunit.

21. A method of making a support for organic synthesis that comprises reacting a magnetic particle with a silane coupling or linking agent and reacting the resulting product with an oligomer subunit.

22. The method of claim 21, wherein the silane coupling or linking agent has a silicon portion and an organic portion, the magnetic particle is convalently attached to said silicon portion and said oligomer subunit is reacted with the organic portion.

23. The method of claim 21, wherein said magnetic particles comprise magnetitie.

24. The method of claim 21, wherein said magnetic particles are non-porous.

25. The method of claim 21, wherein said silane coupling or linking agent has the formula:

space="preserve" listing-type="equation">Si(X).sub.n [R.sub.m (Y).sub.p ].sub.4-n

where Si is the silicon of the organosilane linking or coupling agent;

X is a member of the group of alkyoxy, alkenyloxy, alkynyloxy, alkaryloxy, aryloxy, and halo;

R is a member of the group of alkyl, alkenyl, alkynyl, aryl, alkaryl, or aralkyl;

m is an integer having a value of at least one;

p is an integer with a value of at least one;

Y is a functional group of the group amino, carboxyl or hydroxy, which can react with an oligomer subunit;

n is an integer having a value of 1-3.

26. An oligomer support synthesis having the following structure:

##STR11## wherein M is the support particle, O is the oxygen of the metal oxide or hydroxide, Si is silicon, X is a cleaving group, R is a linking group, Y is a functional group which can react with and bind the oligomer subunit or a group linked thereto, or cross linking agents B and B'"'"' which are chemical bonds or functional groups which react to form covalent bonds with the functional group Y, A is a linking group which bridges the Y groups and is bound to the oligomer subunit to be worked on, either directly or through reactive group C, n is an integer having a value of 1 to 3, m is an integer having a value of at least 1, p is an integer having a value of 1-3, q and r are integers having the value of at least 1, q+r being from 3-7, and s is 0 to 5.

27. The support of claim 26, having the structure:

space="preserve" listing-type="equation">M--O--Si(CH.sub.2).sub.3 --N--(CH.sub.2).sub.2 --N--(CH.sub.2).sub.3 --SI.

28. The support of claim 27, wherein C is --CO(CH₂)₂ CO--;

and Z is a deoxyribonucleotide subunit.