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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MDPI AG
2019-09-01
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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. |
first_indexed | 2024-04-12T21:50:45Z |
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id | doaj.art-6bc3e3a97c074421a5e994cf803fcfd5 |
institution | Directory Open Access Journal |
issn | 1420-3049 |
language | English |
last_indexed | 2024-04-12T21:50:45Z |
publishDate | 2019-09-01 |
publisher | MDPI AG |
record_format | Article |
series | Molecules |
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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