Site Selective Antibody-Oligonucleotide Conjugation via Microbial Transglutaminase

Nucleic Acid Therapeutics (NATs), including siRNAs and AntiSense Oligonucleotides (ASOs), have great potential to drug the undruggable genome. Targeting siRNAs and ASOs to specific cell types of interest has driven dramatic improvement in efficacy and reduction in toxicity. Indeed, conjugation of tr...

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Main Authors: Ian J. Huggins, Carlos A. Medina, Aaron D. Springer, Arjen van den Berg, Satish Jadhav, Xianshu Cui, Steven F. Dowdy
Format: Article
Language:English
Published: MDPI AG 2019-09-01
Series:Molecules
Subjects:
Online Access:https://www.mdpi.com/1420-3049/24/18/3287
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author Ian J. Huggins
Carlos A. Medina
Aaron D. Springer
Arjen van den Berg
Satish Jadhav
Xianshu Cui
Steven F. Dowdy
author_facet Ian J. Huggins
Carlos A. Medina
Aaron D. Springer
Arjen van den Berg
Satish Jadhav
Xianshu Cui
Steven F. Dowdy
author_sort Ian J. Huggins
collection DOAJ
description Nucleic Acid Therapeutics (NATs), including siRNAs and AntiSense Oligonucleotides (ASOs), have great potential to drug the undruggable genome. Targeting siRNAs and ASOs to specific cell types of interest has driven dramatic improvement in efficacy and reduction in toxicity. Indeed, conjugation of tris-GalNAc to siRNAs and ASOs has shown clinical efficacy in targeting diseases driven by liver hepatocytes. However, targeting non-hepatic diseases with oligonucleotide therapeutics has remained problematic for several reasons, including targeting specific cell types and endosomal escape. Monoclonal antibody (mAb) targeting of siRNAs and ASOs has the potential to deliver these drugs to a variety of specific cell and tissue types. However, most conjugation strategies rely on random chemical conjugation through lysine or cysteine residues resulting in conjugate heterogeneity and a distribution of Drug:Antibody Ratios (DAR). To produce homogeneous DAR-2 conjugates with two siRNAs per mAb, we developed a novel two-step conjugation procedure involving microbial transglutaminase (MTGase) tagging of the antibody C-terminus with an azide-functionalized linker peptide that can be subsequently conjugated to dibenzylcyclooctyne (DBCO) bearing oligonucleotides through azide-alkyne cycloaddition. Antibody-siRNA (and ASO) conjugates (ARCs) produced using this strategy are soluble, chemically defined targeted oligonucleotide therapeutics that have the potential to greatly increase the number of targetable cell types.
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spelling doaj.art-6bc3e3a97c074421a5e994cf803fcfd52022-12-22T03:15:29ZengMDPI AGMolecules1420-30492019-09-012418328710.3390/molecules24183287molecules24183287Site Selective Antibody-Oligonucleotide Conjugation via Microbial TransglutaminaseIan J. Huggins0Carlos A. Medina1Aaron D. Springer2Arjen van den Berg3Satish Jadhav4Xianshu Cui5Steven F. Dowdy6Department of Cellular and Molecular Medicine, University of California San Diego, School of Medicine, La Jolla, CA 92093, USADepartment of Cellular and Molecular Medicine, University of California San Diego, School of Medicine, La Jolla, CA 92093, USASorrento Therapeutics, San Diego, CA 92121, USALife Technologies, Thermo Fisher Scientific, Frederick, MD 21703, USADepartment of Cellular and Molecular Medicine, University of California San Diego, School of Medicine, La Jolla, CA 92093, USADepartment of Cellular and Molecular Medicine, University of California San Diego, School of Medicine, La Jolla, CA 92093, USADepartment of Cellular and Molecular Medicine, University of California San Diego, School of Medicine, La Jolla, CA 92093, USANucleic Acid Therapeutics (NATs), including siRNAs and AntiSense Oligonucleotides (ASOs), have great potential to drug the undruggable genome. Targeting siRNAs and ASOs to specific cell types of interest has driven dramatic improvement in efficacy and reduction in toxicity. Indeed, conjugation of tris-GalNAc to siRNAs and ASOs has shown clinical efficacy in targeting diseases driven by liver hepatocytes. However, targeting non-hepatic diseases with oligonucleotide therapeutics has remained problematic for several reasons, including targeting specific cell types and endosomal escape. Monoclonal antibody (mAb) targeting of siRNAs and ASOs has the potential to deliver these drugs to a variety of specific cell and tissue types. However, most conjugation strategies rely on random chemical conjugation through lysine or cysteine residues resulting in conjugate heterogeneity and a distribution of Drug:Antibody Ratios (DAR). To produce homogeneous DAR-2 conjugates with two siRNAs per mAb, we developed a novel two-step conjugation procedure involving microbial transglutaminase (MTGase) tagging of the antibody C-terminus with an azide-functionalized linker peptide that can be subsequently conjugated to dibenzylcyclooctyne (DBCO) bearing oligonucleotides through azide-alkyne cycloaddition. Antibody-siRNA (and ASO) conjugates (ARCs) produced using this strategy are soluble, chemically defined targeted oligonucleotide therapeutics that have the potential to greatly increase the number of targetable cell types.https://www.mdpi.com/1420-3049/24/18/3287oligonucleotide therapeuticssiRNAantisense oligonucleotidesmonoclonal antibodiesantibody-siRNA conjugate (ARC)microbial transglutaminasecopper-less click
spellingShingle Ian J. Huggins
Carlos A. Medina
Aaron D. Springer
Arjen van den Berg
Satish Jadhav
Xianshu Cui
Steven F. Dowdy
Site Selective Antibody-Oligonucleotide Conjugation via Microbial Transglutaminase
Molecules
oligonucleotide therapeutics
siRNA
antisense oligonucleotides
monoclonal antibodies
antibody-siRNA conjugate (ARC)
microbial transglutaminase
copper-less click
title Site Selective Antibody-Oligonucleotide Conjugation via Microbial Transglutaminase
title_full Site Selective Antibody-Oligonucleotide Conjugation via Microbial Transglutaminase
title_fullStr Site Selective Antibody-Oligonucleotide Conjugation via Microbial Transglutaminase
title_full_unstemmed Site Selective Antibody-Oligonucleotide Conjugation via Microbial Transglutaminase
title_short Site Selective Antibody-Oligonucleotide Conjugation via Microbial Transglutaminase
title_sort site selective antibody oligonucleotide conjugation via microbial transglutaminase
topic oligonucleotide therapeutics
siRNA
antisense oligonucleotides
monoclonal antibodies
antibody-siRNA conjugate (ARC)
microbial transglutaminase
copper-less click
url https://www.mdpi.com/1420-3049/24/18/3287
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