Work

Protocol to generate, purify, and analyze antibody-oligonucleotide conjugates from off-the-shelf antibodies

Year: 2024

Type: article

Abstract: Antibody-oligonucleotide conjugates (AOCs) are a fast-expanding modality for targeted delivery of therapeutic oligonucleotides to tissues. Here, we present a protocol to generate, purify, and analyze AOCs from off-the-shelf antibodies. We describe steps to conjugate single/double-stranded oligonucleotides bearing amine handles to linkers and, then, to antibodies using well-established chemistry. In addition, we provide details regarding the purification techniques and analytical methods suitable for AOC. This protocol can be applied for several purposes where AOC is a modality of interest. For complete details on the use and execution of this protocol, please refer to Rady et al. 1 • Straightforward method to conjugate amino-modified oligonucleotides to linkers • Steps for hybridizing linker-oligonucleotide to its complementary strand • Fast and reproducible lysine conjugation of azide groups to off-the-shelf antibodies • State-of-the-art purification and analysis methods of antibody-siRNA conjugates Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. Antibody-oligonucleotide conjugates (AOCs) are a fast-expanding modality for targeted delivery of therapeutic oligonucleotides to tissues. Here, we present a protocol to generate, purify, and analyze AOCs from off-the-shelf antibodies. We describe steps to conjugate single/double-stranded oligonucleotides bearing amine handles to linkers and, then, to antibodies using well-established chemistry. In addition, we provide details regarding the purification techniques and analytical methods suitable for AOC. This protocol can be applied for several purposes where AOC is a modality of interest.

Source: STAR Protocols

DOI: 10.1016/j.xpro.2024.103329