A Platform for Site‐Specific DNA‐Antibody Bioconjugation by Using Benzoylacrylic‐Labelled Oligonucleotides

Abstract

Many bioconjugation strategies for DNAo ligonucleotides and antibodies suffer limitations,s uch as sitespecificity,s toichiometry and hydrolytic instability of the conjugates,w hichm akes them unsuitable for biological applications.H ere,w er eport an ew platform for the preparation of DNA-antibody bioconjugates with as imple benzoylacrylic acid pentafluorophenyl ester reagent. Benzoylacryliclabelled oligonucleotides prepared with this reagent can be sitespecifically conjugated to ar ange of proteins and antibodies through accessible cysteine residues.T he homogeneity of the prepared DNA-antibody bioconjugates was confirmed by an ew LC-MS protocol and the bioconjugate probes were used in fluorescence or super-resolution microscopyc ell imaging experiments.T his work demonstrates the versatility and robustness of our bioconjugation protocol that gives sitespecific,w ell-defined and plasma-stable DNA-antibody bioconjugates for biological applications.

Bioconjugation via Hetero‐Selective Clamping of Two Different Amines withortho‐Phthalaldehyde

“…The key to an optimal three‐component bioconjugation with two nucleophiles is control of hetero‐crosslinking selectivity. Bifunctional linkers such as those with a N‐hydroxysuccinimide (NHS), maleimide, disulfide, or α‐haloacetamide group have been widely used to crosslink amines to the thiol group on biomolecules [18–22] . However, moderately reactive groups, e.g.…”