Methods for the development of vaccines based on oligosaccharide-oligonucleotide conjugates

摘要

Described herein are oligosaccharide-oligonucleotide conjugates useful as vaccines against one or more human or veterinary therapeutic indications, and methods of synthesizing and identifying them. The conjugates may be identified using non-human antibodies as binding targets, thereby expanding the power and scope of the invention. Efficacious conjugates may be identified through an iterative.

主权项

1. A method, comprising the steps of:
(a) combining an oligonucleotide, a first DNA polymerase, and a plurality of deoxyribonucleotide triphosphates,
whereinthe oligonucleotide comprises a first primer binding site on the 5′ end, a randomized region, and a stem-loop region;the randomized region is located between the first primer binding site and the stem-loop region;the stem-loop region comprises a second primer binding site;at least one of the deoxyribonucleotide triphosphates comprises a reactive substituent; andthe reactive substituent is ethynyl, thereby forming an extended oligonucleotide comprising an original strand and an extended strand, wherein the extended strand comprises at least one reactive substituent; (b) combining a plurality of modifying compounds and the extended oligonucleotide under reaction conditions, wherein the modifying compound is a compound of formula III: wherein A represents a branched or unbranched oligosaccharide consisting of about 3 to about 15 saccharide moieties, thereby forming a modified extended oligonucleotide comprising the original strand and a modified extended strand; and (c) combining a primer complementary to the second primer binding site, a second DNA polymerase, the modified extended oligonucleotide, and a plurality of deoxyribonucleotide triphosphates thereby creating a duplex with the original strand and displacing the modified extended strand.