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Cross-Reactive Polyclonal Antibodies Raised Against GalNAc-Conjugated siRNA Recognize Mostly the GalNAc Moiety

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Abstract

Small interfering RNA (siRNA) is gaining momentum as a therapeutic modality with six approved products. Since siRNA has the potential to elicit undesired immune responses in patients, immunogenicity assessment is required during clinical development by regulatory authorities. In this study, anti-siRNA polyclonal antibodies were generated through animal immunization. These cross-reactive polyclonal antibodies recognized mostly the N-acetylgalactosamine (GalNAc) moiety with a small fraction against sequence-independent epitopes. We demonstrate that the polyclonal antibodies can be utilized as immunogenicity assay positive controls for the same class of GalNAc-conjugated siRNAs. In addition, anti-GalNAc mAbs showed desired sensitivity and drug tolerance, supporting their use as alternative surrogate positive controls. These findings can guide positive control selection and immunogenicity assay development for GalNAc-conjugated siRNAs and other oligonucleotide therapeutics.

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Acknowledgements

The authors would like to thank Drs. Martin McGrath and Nicoletta Bivi for their help and discussion on reagent generation. The authors would like to thank Drs. Kelly Hainline, Yuewei Qian, and Robert Siegel for reviewing the manuscript.

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Authors and Affiliations

  1. Lilly Research Laboratories, Eli Lilly and Company, Indianapolis, 46285, Indiana, USA

    Kimberly K. Ballman, Victoria L. Peek, John H. Sloan, Jingling Li, Robert J. Konrad & Yi Wen

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  1. Kimberly K. Ballman
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  2. Victoria L. Peek
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  3. John H. Sloan
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  4. Jingling Li
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  5. Robert J. Konrad
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  6. Yi Wen
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Contributions

K. B. and Y. W. designed the study. K. B., V. P., J. S., and J. L. performed the experiments and generated the data or reagents. K. B. and Y. W. analyzed the data. R. K. provided supervision and edited the manuscript. Y. W. supervised the study and wrote the manuscript. All authors reviewed and approved the manuscript.

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Correspondence to Yi Wen.

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All authors are employees of Eli Lilly and Company and own company stocks.

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Ballman, K.K., Peek, V.L., Sloan, J.H. et al. Cross-Reactive Polyclonal Antibodies Raised Against GalNAc-Conjugated siRNA Recognize Mostly the GalNAc Moiety. AAPS J 26, 41 (2024). https://doi.org/10.1208/s12248-024-00914-w

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