Purification of oligomers using dual-end selection
AssigneeBayer Aktiengesellschaft
InventorsThomas Horn, Michael S. Urdea
Life of this patent
Estimating the term…
Abstract
Oligomers are prepared substantially free of error sequences by sequentially adding monomers to a growing chain bound to a support through a first selectably cleavable linkage, a first capture moiety and a second selectably cleavable linkage. At the completion of monomer addition, the completed oligomer is cleaved from the support to reveal the first capture moiety and purified by virtue of the presence of a second capture moiety, e.g., a terminal blocking group, and the first capture moiety. A support-bound oligomer having the structural formula (I) S—[X1]n1—SC1—CP2—[X2]n2—SC3—T1—X—T2—SC2—CP1 (I) is also provided wherein T1, T2, X1, X2, n1, n2, SC1, SC2, SC3, CP1 and CP2 are as defined herein.
Claims16
A method for preparing a purified oligomer segment of interest, comprising:
- (a) providing a support-bound oligomer selected from the group consisting of oligonucleotides and oligopeptides having a first selectably cleavable linkage, a second selectably cleavable linkage and a third selectably cleavable linkage, wherein the oligomer segment of interest is flanked by the second and third selectably cleavable linkages, and further wherein a first capture moiety is present at the free terminus of the oligomer and a second capture moiety is present between the first and third selectably cleavable linkages;
- (b) cleaving the first selectably cleavable linkage to release the oligomer from the support;
- (c) incubating the released oligomer with a first capture medium that selectively retains the released oligomer by binding to the first capture moiety, to form a first capture medium-oligomer complex;
- (d) cleaving the second selectably cleavable linkage;
- (e) incubating the oligomeric product of step (d) with a second capture medium that selectively binds to the second capture moiety, to form a second capture medium-oligomer complex;
- and
- (f) cleaving the third selectably cleavable linkage to provide the oligomer segment of interest in purified form.
A support-bound oligomer selected from the group consisting of oligonucleotides and oligopeptides having the structural formula (I)
- S—[X1]n1—SC1—CP2—[X2]n2—SC3—T1—X—T2—SC2—CP1 (I)
- wherein:
- T1 and T2 represent first and second oligomer termini, respectively;
- S represents a solid support;
- X represents an oligomer segment of interest;
- X1 and X2 are monomers or oligomeric segments;
- n1 and n2 are independently zero or 1;
- SC1 represents a first selectably cleavable linkage;
- SC2 represents a second selectably cleavable linkage;
- SC3 represents a third selectably cleavable linkage;
- CP1 represents a first capture moiety;
- and
- CP2 represents a second capture moiety.
A method for preparing a purified oligomer segment of interest, comprising:
- (a) providing a support-bound oligomer selected from the group consisting of oligonucleotides and oligopeptides having the structural formula (I)
- S—[X1]n1—SC1—CP2—[X2]n2—SC3—T1—X—T2—SC2—CP1 (I)
- wherein:
- T1 and T2 represent first and second oligomer termini, respectively;
- S represents a solid support;
- X represents an oligomer segment of interest;
- X1 and X2 are monomers or oligomeric segments;
- n1 and n2 are independently zero or 1;
- SC1 represents a first selectably cleavable linkage;
- SC2 represents a second selectably cleavable linkage;
- SC3 represents a third selectably cleavable linkage;
- CP1 represents a first capture moiety;
- and
- CP2 represents a second capture moiety;
- (b) cleaving the first selectably cleavable linkage to release the oligomer from the support;
- (c) incubating the released oligomer with a first capture medium that selectively retains the released oligomer by binding to the first capture moiety, to form a first capture medium-oligomer complex;
- (d) cleaving the second selectably cleavable linkage;
- (e) incubating the oligomeric product of step (d) with a second capture medium that selectively binds to the second capture moiety, to form a second capture medium-oligomer complex;
- and
- (f) cleaving the third selectably cleavable linkage to provide the oligomer segment of interest in purified form.
Dependencies are inferred from the claim text.