Polymer reversal on solid surfaces

• US 5,550,215 A
• Filed: 11/28/1994
• Issued: 08/27/1996
• Est. Priority Date: 11/22/1991
• Status: Expired due to Term

• Current Assignee: Affymetrix, Inc. (Thermo Fisher Scientific Incorporated)
• Original Assignee: Affymetrix, Inc. (Thermo Fisher Scientific Incorporated)
• Inventors: Holmes, Christopher P.
• Primary Examiner(s): Russel, Jeffrey E.
• Application Number:
  US08/351,058
• Time in Patent Office: 638 Days
• Field of Search: 536/25.3, 536/25.31, 536/124, 536/126, 530/317, 530/318, 530/319, 530/320, 530/321, 530/332, 530/334, 530/345, 530/335, 530/336, 530/337, 525/54.11, 525/54.2
• US Class Current: 506/30
• CPC Class Codes: B01J 2219/00432 Photolithographic masks
• CPC Class Codes: B01J 2219/00527 Sheets
• CPC Class Codes: B01J 2219/00529 DNA chips
• CPC Class Codes: B01J 2219/00585 Parallel processes
• CPC Class Codes: B01J 2219/0059 Sequential processes
• CPC Class Codes: B01J 2219/00596 Solid-phase processes
• CPC Class Codes: B01J 2219/00605 the compounds being directly bound or immobilised to solid supports
• CPC Class Codes: B01J 2219/00608 DNA chips
• CPC Class Codes: B01J 2219/0061 The surface being organic
• CPC Class Codes: B01J 2219/00612 the surface being inorganic
• CPC Class Codes: B01J 2219/00626 Covalent
• CPC Class Codes: B01J 2219/00659 Two-dimensional arrays
• CPC Class Codes: B01J 2219/00711 Light-directed synthesis
• CPC Class Codes: B01J 2219/00722 Nucleotides
• CPC Class Codes: B01J 2219/00725 Peptides
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C07B 2200/11 Compounds covalently bound to a solid support
• 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 1/04 on carriers C07K1/003, C07K1/006 take precedence
• CPC Class Codes: C07K 1/042 characterised by the nature of the carrier
• CPC Class Codes: C40B 40/06 : Libraries containing nucleotides or polynucleotides, or derivatives thereof
• CPC Class Codes: C40B 40/10 : Libraries containing peptides or polypeptides, or derivatives thereof
• CPC Class Codes: C40B 60/14 : for creating libraries
• CPC Class Codes: G03F 7/00 : Photomechanical, e.g. photolithographic, production of textured or patterned surfaces, e.g. printing surfaces; Materials therefor, e.g. comprising photoresists; Apparatus specially adapted therefor using photoresist structures for special production processes, see the relevant places, e.g. B44C, H01L, e.g. H01L21/00, H05K
• CPC Class Codes: Y02P 20/55 : Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups

Abstract

A method for cyclization and reversal of the polarity of polymers on a substrate. The method provides for the formation of a polymer on a substrate (2) with a tether molecule (4). Through unmasking of a protective group (PG₂) a cyclic polymer (6) is formed. Through cleavage of an appropriate bond, a polarity reversed polymer (8) is formed. The method finds particular application in the formation of, for example, peptides and oligonucleotides.

Claims

1. A method of synthesizing a polymer on a substrate comprising:

a) on a surface of said substrate attaching a molecule having the general formula ##STR17## where;

X is a first tether reactive site selected from the group consisting of O, NH, S, CO₂, S--(CH₂)_n O and NHCO(CH₂)_n CO₂, where n is one to ten,

Y is a second tether reactive site selected from the group consisting of;

1) O, NH, S, CO₂, S--(CH₂)_n O, and NHCO(CH₂)_n CO₂ functionality which functionality is blocked with a second protective group; and

2) OH, NH₂, SH, CO₂ H, S--(CH₂)_n OH and NHCO(CH₂)_n CO₂ H functionality, where n is one to ten,

T is a tether molecule, and

PG is a first protective group;

wherein the molecule is attached to the surface via a covalent bond between T and the surface;

b) removing any such second protective group employed on the second tether site Y while retaining the first protective group;

(c) coupling a polymer, said polymer having a first polymer reactive site and a second polymer reactive site, to said tether reactive site Y, wherein a bond is formed between said second polymer reactive site and said tether reactive site Y;

(d) removing said first protective group PG;

(e) coupling said first polymer reactive site to said tether at said first tether reactive site X; and

(f) cleaving said bond between Y and the polymer whereby the polarity of the said polymer is reversed

wherein said polymer is a linear, cyclic or branched chain polymer selected from the group consisting of peptides, nucleic acids and polysaccharides.

2. A method recited in claim 1 wherein said second tether reactive site, Y, is blocked with a second protective group and further wherein the procedure of coupling a polymer to said tether reactive site Y comprises:

i) removing said second protective group from said tether reactive site Y;

ii) coupling a first monomer to said tether reactive site Y, said first monomer having a monomer reactive site protected with a first monomer protective group;

iii) repeating said steps of removing a protective group and coupling a monomer to form a polymer at said tether reactive site Y, wherein said polymer has a first polymer reactive site and a second polymer reactive site and wherein said second polymer reactive site is coupled to said tether at said second tether reactive site.

4. The method as recited in claim 2 wherein said first protective group is NVOC and said second protective group is selected from the group consisting of FMOC, allyl, and DMT.

5. The method as recited in claim 2 wherein said first protective group is selected from the group consisting of FMOC and DMT and said second protective group is NVOC.

6. The method as recited in claim 1 wherein said step of cleaving comprises the step of exposing said polymer to an acid.

7. The method as recited in claim 1 wherein said step of cleaving comprises the step of exposing said polymer to light.

9. A method as recited in claim 2 wherein said molecule having the formula ##STR20## is a molecule having the general formula ##STR21## where:

X and Y are first and second reactive sites, respectively;

PG₁ and PG₂ are first and second protecting groups, respectively;

R is selected from the group consisting of hydrogen, methyl, phenyl, substituted phenyl, and bridged phenyl; and

R'"'"' is selected from the group consisting of methyl, phenyl, substituted phenyl, and bridged phenyl.

10. A method as recited in claim 2 wherein said molecule having the general formula:

##STR22## is a molecule having the general formula ##STR23## where;

X and Y are first and second reactive sites, respectively;

PG₁ and PG₂ are first and second protecting groups, respectively; and

R and R'"'"' are independently selected from the group consisting of hydrogen, methyl, phenyl, substituted phenyl, and bridged phenyl.

11. A method as recited in claim 2 wherein said molecule having the general formula:

##STR24## is a molecule having the general formula ##STR25## where;

X and Y are first and second reactive sites, respectively;

PG₁ and PG₂ are first and second protecting groups, respectively; and

R and R'"'"' are independently selected from the group consisting of hydrogen, methyl, phenyl, substituted phenyl, and bridged phenyl.

12. A method as recited in claim 2 wherein said molecule having the general formula:

##STR26## is a molecule having the general formula ##STR27## where;

X and Y are first and second reactive sites, respectively;

PG₁ and PG₂ are first and second protecting groups, respectively; and

R and R'"'"' are independently selected from the group consisting of hydrogen, methyl, phenyl, substituted phenyl, and bridged phenyl.

13. A method as recited in claim 2 wherein said molecule having the general formula:

##STR28## is a molecule having the general formula ##STR29## where;

X and Y are first and second reactive sites, respectively;

PG₁ and PG₂ are first and second protecting groups, respectively; and

R and R'"'"' are independently selected from the group consisting of hydrogen, methyl, phenyl, substituted phenyl, and bridged phenyl.

14. A method as recited in claim 2 wherein said first protective group PG is acid-labile and said second protective group is selected from the group consisting of base-labile and photoreactive protective groups.

15. A method as recited in claim 2 wherein said first protective group PG is base-labile and said second protective group is selected from the group consisting of acid-labile and photoreactive protective groups.

16. A method as recited in claim 2 wherein said first protective group PG is a photoreactive protective group and said second protective group is selected from the group consisting of acid-labile and base-labile protective groups.

3. A method of synthesizing a polymer on a substrate comprising:

a) on a surface of said substrate attaching a molecule having the general formula ##STR18## wherein;

X is a first tether reactive site selected from the group consisting of O, NH, S, CO₂, S--(CH₂)_n O, and NHCO(CH₂)_n CO₂, where n is one to ten,

Y is a second tether reactive site selected from the group consisting of;

1) O, NH, S, CO₂, S--(CH₂)_n O, and NHCO(CH₂)_n CO₂ functionality which functionality is blocked with a second protective group; and

2) OH, NH₂, SH, CO₂ H, S--(CH₂)_n OH and NHCO(CH₂)_n CO₂ H functionality, where n is one to ten.

T is a tether molecule, and

PG is a first protective group;

wherein the molecule is attached to the surface via a covalent bond between T and the surface;

(b) removing any such second protective group employed on the second tether site Y while retaining the first protective group;

(c) coupling a polymer, said polymer having a first polymer reactive site and a second polymer reactive site, to said tether reactive site Y, wherein a bond is formed between said second polymer reactive site and said tether reactive site Y;

(d) removing said first protective group PC;

(e) coupling said first polymer reactive site to said tether at said first tether reactive site X; and

(f) cleaving said bond between Y and the polymer whereby a polarity of the said polymer is reversed

wherein said polymer is a linear, cyclic or branched chain peptide.

8. A method as recited in claim 3 wherein said molecule having the formula ##STR19## is an amino acid containing a protecting group PG on either the amino moiety or the carboxyl moiety.