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Most synthetic schemes to generate these hybrid molecules require expensive reagents, significant quantities of input antibody, and multistep purification routes; thus limiting widespread application. Herein a facile and robust conjugation strategy is reported that involves \u201cplug\u2010and\u2010play\u201d antibody conjugation with succinimidyl\u2010functionalized oligonucleotides, which are high yielding and compatible for use directly after buffer exchange. The succinimidyl\u2010linked oligonucleotides are synthesized with 5\u2032\u2010amine\u2010modified oligonucleotides and disuccinimidyl suberate (DSS), both of which are inexpensive and commercially available. Direct incubation of the resulting stable succinimidyl\u2013 oligonucleotide conjugates with commercial antibodies yields conjugates ready for use after benchtop buffer exchange. It is demonstrated that the resulting oligonucleotide\u2013antibody and oligonucleotide\u2013streptavidin conjugates retain potent and specific binding in activity\u2010dependent proximity ligation imaging, and proximity ligation\u2010mediated qPCR detection of endogenous proteins in native cellular contexts down to picogram levels of whole proteome. This DSS conjugation strategy should be widely applicable in the synthesis of protein\u2013oligonucleotide conjugates.<\/jats:p>","DOI":"10.1002\/cbic.201900027","type":"journal-article","created":{"date-parts":[[2019,2,14]],"date-time":"2019-02-14T21:46:14Z","timestamp":1550180774000},"page":"1599-1605","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":27,"title":["A Concise, Modular Antibody\u2013Oligonucleotide Conjugation Strategy Based on Disuccinimidyl Ester Activation Chemistry"],"prefix":"10.1002","volume":"20","author":[{"given":"Gang","family":"Li","sequence":"first","affiliation":[{"name":"Department of Chemistry University of Chicago  929 E. 57th Street Chicago IL 60637 USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2043-7838","authenticated-orcid":false,"given":"Raymond E.","family":"Moellering","sequence":"additional","affiliation":[{"name":"Department of 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