Membranes with bound oligonucleotides and peptides
AssigneeMillipore Corporation
InventorsHubert Koester, James M. Coull
Life of this patent
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Abstract
A method is provided for synthesizing oligonucleotides and peptides directly onto a membrane. The method provides a means for generating membrane affinity supports. A modified membrane for the method of direct synthesis is also provided.
Claims42
A modified membrane comprising a polymeric membrane having a protected nucleoside or amino acid linked thereto.
A modified membrane represented by the formula:
- P--X--Y--N--Z--S.sup.W
- wherein
- P is a polymeric membrane;
- X is a functional group on the membrane;
- Y--N--Z is a linker wherein N is a spacer molecule and Y and Z are the same or different functional groups, the linker being bound to the membrane through its functional group Y to the functional group X;
- and
- S.sup.W is a protected nucleoside or amino acid, S.sup.W being bound to the linker through the functional group Z of the linker.
A method of synthesizing an oligonucleotide comprising the steps of:
- a. providing a modified membrane represented by the formula:
- P--X--Y--N--Z--S.sup.W
- wherein
- P is a polymeric membrane;
- X is a functional group on the membrane;
- Y--N--Z is a linker wherein N is a spacer molecule and Y and Z are the same or different functional groups, the linker being bound to the membrane through the functional group X;
- and
- S.sup.W is a protected nucleoside, S.sup.W being bound to the linker through the functional group Z of the linker;
- b. coupling a nucleoside phosphoramidite to the nucleoside S.sup.W to produce a membrane-bound nucleoside-nucleotide having a phosphite triester linkage;
- c. oxidizing the phosphite triester to form a phosphate triester linkage;
- and
- d. sequentially coupling additional nucleoside phosphoramidites to the membrane-bound nucleoside-nucleotide, and after each coupling step, oxidizing the resulting phosphite triester linkage to a phosphate triester to produce a membrane-bound polynucleotide.
A method of synthesizing a peptide, comprising the steps of:
- a. providing a modified membrane of the formula
- P--X--Y--N--Z--S.sup.W
- wherein
- P is a polymeric membrane;
- X is a functional group on the membrane;
- Y--N--Z is a linker wherein N is a spacer molecule and Y and Z are the same or differential functional groups, the linker being bound to the membrane through the functional group X;
- and
- S.sup.W is a amino acid, S.sup.W being bound to the linker through the functional group Z of the linker;
- and
- b. sequentially coupling amino acids to S.sup.W to produce a membrane-bound peptide.
A membrane having an oligonucleotide attached through a terminus of the oligonucleotide.
A membrane having an peptide attached through a terminus of the peptide.
A membrane having an attached oligonucleotide or peptide represented by the formula:
- P--X--Y--N--Z--(--S.sup.W).sub.n
- wherein
- P is a polymeric membrane;
- X is a functional group on the membrane;
- Y--N--Z is a linker wherein N is a spacer molecule and Y and Z are the same or different functional groups, the linker being bound to the membrane through the functional group X;
- and
- (S.sup.W).sub.n represents an oligonucleotide or peptide comprised of protected or partly protected nucleotides or amino acids, wherein n is the number of nucleotide or amino acid units of the oligonucleotide,
- (S.sup.W).sub.n being bound to the linker through the functional group Z of the linker.
A membrane having an attached oligonucleotide or peptide represented by the formula:
- P--X--Y--N--Z--(--S).sub.n
- wherein
- P is a polymeric membrane;
- X is a functional group on the membrane;
- Y--N--Z is a linker wherein N is a spacer molecule and Y and Z are the same or different functional groups, the linker being bound to the membrane through the functional group X;
- and
- (S).sub.n represents an oligonucleotide or peptide, wherein n is the number of nucleotide or amino acid units of the oligonucleotide or peptide, (S).sub.n being bound to the linker through the functional group Z of the linker.
A modified membrane of the formula:
- ##STR8## wherein P is a polyvinylidene difluoride;
- m is 1 through 6;
- B.sup.W is a nucleoside base having a protecting group for protection of exocylic amino groups;
- and
- DMT is the 4,4-dimethoxytrityl protecting group.
A modified membrane of the formula:
- ##STR9## wherein P is a membrane comprising polypropylene coated with hydroxyporpylacrylate;
- n=1-20;
- Nle is norleucine;
- U.sup.W is an amino acid side chain;
- and
- Fmoc is a fluorenylmethoxycarbonyl protecting group.
Dependencies are inferred from the claim text.