Synthesis of peptide-oligonucleotide conjugates using a heterobifunctional crosslinker.
Peptide-oligonucleotide conjugates (POCs) are molecular chimeras composed of a nucleic acid moiety covalently attached to a polypeptide moiety. POCs have been used in numerous applications from therapeutics to nanotechnology, and most recently as combinatorial agents in the assembly of bivalent protein affinity reagents. This unit describes the synthesis and purification of POC molecules using the heterobifunctional crosslinking reagent succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC), which enables amine-modified oligonucleotides to become covalently linked to cysteine-modified polypeptides. This solution-based protocol consists of a two-step synthesis followed by a single purification step.
Williams
Berea A R
BA
Department of Chemistry and Biochemistry, Arizona State University, Tempe, AZ, USA.
Chaput
John C
JC
D002851:Chromatography, High Pressure Liquid; D003432:Cross-Linking Reagents; D024202:Electrophoretic Mobility Shift Assay; D008301:Maleimides / Q000737:chemistry*; D008958:Models, Molecular; D009841:Oligonucleotides / Q000138:chemical synthesis* / Q000737:chemistry; D010455:Peptides / Q000138:chemical synthesis* / Q000737:chemistry; D011506:Proteins / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D003432:Cross-Linking Reagents; D008301:Maleimides; D009841:Oligonucleotides; D010455:Peptides; D011506:Proteins
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