posted on 2019-06-27, 00:00authored byMikkel
B. Skovsgaard, Michael R. Mortensen, Johan Palmfeldt, Kurt V. Gothelf
A method
for aptamer directed conjugation of DNA to therapeutic
antibodies has been developed. In the method, an antibody selective
aptamer binds to a specific site in the constant domain of human IgG1
antibodies and is used for both templated and direct conjugation to
the antibodies. Through optimization of the design and reaction conditions,
the antibody–DNA conjugates could be produced efficiently using
equal stoichiometry of protein and DNA. Three different antibodies
were evaluated, and the conjugates were characterized by anion exchange
chromatography and SDS-PAGE. The conjugation sites for one of the
antibodies were determined by MS/MS analysis of the digested conjugate.
The antibody–DNA conjugate was also tested for receptor binding
on cell surfaces showing retained binding.