A Robust and Automated Platform for Charge Detection Mass Spectrometry of Megadalton Biotherapeutics

Samuel E. Janisse; Ryan T. Fellers; Shannon A. Raab; Michael P. Goodwin; Kyle P. Bowen; Zhirui J. Lian; Kenneth R. Durbin; Michael W. Senko; Philip D Compton; Jared O. Kafader; Neil L Kelleher

doi:10.1021/acs.analchem.4c06397

A Robust and Automated Platform for Charge Detection Mass Spectrometry of Megadalton Biotherapeutics

Samuel E. Janisse, Ryan T. Fellers, Shannon A. Raab, Michael P. Goodwin, Kyle P. Bowen, Zhirui J. Lian, Kenneth R. Durbin, Michael W. Senko, Philip D Compton, Jared O. Kafader, Neil L Kelleher^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

Abstract

Gene therapies based on adeno-associated viruses are an emerging area with high potential to improve human health. Current quality control techniques to assess contaminates and byproducts from the adeno-associated virus (AAV) production pipelines are lacking in robustness and throughput. To address these limitations, we coupled an automated microfluidic device called SampleStream with Orbitrap-based charge detection mass spectrometry (SS-CDMS). We demonstrate that the SS-CDMS workflow performs AAV analysis in under 15 min per sample in a completely autonomous manner. The SS-CDMS workflow enables rapid assessment of key quality control attributes (CQAs), such as of molecular weight and content ratio of AAV formulations with a small sample requirement (<2 × 10⁹ capsids) without being limited by sample concentration. Additionally, this work shows the potential for the SS-CDMS workflow to be implemented at various stages of the production pipeline through effective sample clean up from more complex AAV matrices such as cell culture media.

Original language	English (US)
Pages (from-to)	4549-4555
Number of pages	7
Journal	Analytical Chemistry
Volume	97
Issue number	8
DOIs	https://doi.org/10.1021/acs.analchem.4c06397
State	Published - Mar 4 2025

Funding

This work was supported by National Institutes of Health P41 GM108569 (N.L.K.), the NIH Office of Director award S10 OD025194 (P.D.C.), and the National Institutes of Health P30 CA060553 (awarded to the Robert H. Lurie Comprehensive Center).

ASJC Scopus subject areas

Analytical Chemistry

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title = "A Robust and Automated Platform for Charge Detection Mass Spectrometry of Megadalton Biotherapeutics",

abstract = "Gene therapies based on adeno-associated viruses are an emerging area with high potential to improve human health. Current quality control techniques to assess contaminates and byproducts from the adeno-associated virus (AAV) production pipelines are lacking in robustness and throughput. To address these limitations, we coupled an automated microfluidic device called SampleStream with Orbitrap-based charge detection mass spectrometry (SS-CDMS). We demonstrate that the SS-CDMS workflow performs AAV analysis in under 15 min per sample in a completely autonomous manner. The SS-CDMS workflow enables rapid assessment of key quality control attributes (CQAs), such as of molecular weight and content ratio of AAV formulations with a small sample requirement (<2 × 109 capsids) without being limited by sample concentration. Additionally, this work shows the potential for the SS-CDMS workflow to be implemented at various stages of the production pipeline through effective sample clean up from more complex AAV matrices such as cell culture media.",

author = "Janisse, {Samuel E.} and Fellers, {Ryan T.} and Raab, {Shannon A.} and Goodwin, {Michael P.} and Bowen, {Kyle P.} and Lian, {Zhirui J.} and Durbin, {Kenneth R.} and Senko, {Michael W.} and Compton, {Philip D} and Kafader, {Jared O.} and Kelleher, {Neil L}",

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doi = "10.1021/acs.analchem.4c06397",

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AU - Janisse, Samuel E.

AU - Fellers, Ryan T.

AU - Raab, Shannon A.

AU - Goodwin, Michael P.

AU - Bowen, Kyle P.

AU - Lian, Zhirui J.

AU - Durbin, Kenneth R.

AU - Senko, Michael W.

AU - Compton, Philip D

AU - Kafader, Jared O.

AU - Kelleher, Neil L

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N2 - Gene therapies based on adeno-associated viruses are an emerging area with high potential to improve human health. Current quality control techniques to assess contaminates and byproducts from the adeno-associated virus (AAV) production pipelines are lacking in robustness and throughput. To address these limitations, we coupled an automated microfluidic device called SampleStream with Orbitrap-based charge detection mass spectrometry (SS-CDMS). We demonstrate that the SS-CDMS workflow performs AAV analysis in under 15 min per sample in a completely autonomous manner. The SS-CDMS workflow enables rapid assessment of key quality control attributes (CQAs), such as of molecular weight and content ratio of AAV formulations with a small sample requirement (<2 × 109 capsids) without being limited by sample concentration. Additionally, this work shows the potential for the SS-CDMS workflow to be implemented at various stages of the production pipeline through effective sample clean up from more complex AAV matrices such as cell culture media.

AB - Gene therapies based on adeno-associated viruses are an emerging area with high potential to improve human health. Current quality control techniques to assess contaminates and byproducts from the adeno-associated virus (AAV) production pipelines are lacking in robustness and throughput. To address these limitations, we coupled an automated microfluidic device called SampleStream with Orbitrap-based charge detection mass spectrometry (SS-CDMS). We demonstrate that the SS-CDMS workflow performs AAV analysis in under 15 min per sample in a completely autonomous manner. The SS-CDMS workflow enables rapid assessment of key quality control attributes (CQAs), such as of molecular weight and content ratio of AAV formulations with a small sample requirement (<2 × 109 capsids) without being limited by sample concentration. Additionally, this work shows the potential for the SS-CDMS workflow to be implemented at various stages of the production pipeline through effective sample clean up from more complex AAV matrices such as cell culture media.

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A Robust and Automated Platform for Charge Detection Mass Spectrometry of Megadalton Biotherapeutics

Abstract

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ASJC Scopus subject areas

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