Nanoscale Mapping of Recombinant Viral Proteins: From Cells to Virus-Like Particles

Maria Arista-Romero; Pietro Delcanale; Silvia Pujals; Lorenzo Albertazzi

doi:10.1021/acsphotonics.1c01154

Nanoscale Mapping of Recombinant Viral Proteins: From Cells to Virus-Like Particles

Maria Arista-Romero, Pietro Delcanale, Silvia Pujals (Corresponding author), Lorenzo Albertazzi (Corresponding author)

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

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Abstract

Influenza recombinant proteins and virus-like particles (VLPs) play an important role in vaccine development (e.g., CadiFlu-S). However, their production from mammalian cells suffers from low yields and lack of control of the final VLPs. To improve these issues, characterization techniques able to visualize and quantify the different steps of the process are needed. Fluorescence microscopy represents a powerful tool able to image multiple protein targets; however, its limited resolution hinders the study of viral constructs. Here, we propose the use of super-resolution microscopy and in particular of DNA-point accumulation for imaging in nanoscale topography (DNA-PAINT) microscopy as a characterization method for recombinant viral proteins on both cells and VLPs. We were able to quantify the amount of the three main influenza proteins (hemagglutinin (HA), neuraminidase (NA), and ion channel matrix protein 2 (M2)) per cell and per VLP with nanometer resolution and single-molecule sensitivity, proving that DNA-PAINT is a powerful technique to characterize recombinant viral constructs.

Original language	English
Pages (from-to)	101–109
Number of pages	9
Journal	ACS Photonics
Volume	9
Issue number	1
DOIs	https://doi.org/10.1021/acsphotonics.1c01154
Publication status	Published - 19 Jan 2022

Bibliographical note

Funding Information:
The authors would like to acknowledge the groups of Dr. Nico Callewaert and Dr. Xavier Saelens from VIB-UGENT Center for Medical Biotechnology for providing the influenza plasmids used in this paper (PHW2000 HA, NA, and M). M.A.-R. is grateful to the Spanish Ministry of Universities and the European Social Fund for the funding of his research activity by the research project (FPI16-SEV-2014-0425-16-1) (Ayudas para contratos predoctorales para la formación de doctores 2016). S.P. and L.A acknowledge the financial support by the Spanish Ministry of Science and Innovation (PID2019-109450RB-I00/AEI/10.13039/501100011033), European Research Council/Horizon 2020 (ERC-StG-757397), “la Caixa” Foundation (ID 100010434), and the Generalitat de Catalunya (through the CERCA program and 2017 SGR 01536).

Keywords

DNA-PAINT
recombinant proteins
single-molecule localization microscopy
single-particle analysis
super-resolution microscopy
virus-like particles

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1021/acsphotonics.1c01154

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Cite this

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title = "Nanoscale Mapping of Recombinant Viral Proteins: From Cells to Virus-Like Particles",

abstract = "Influenza recombinant proteins and virus-like particles (VLPs) play an important role in vaccine development (e.g., CadiFlu-S). However, their production from mammalian cells suffers from low yields and lack of control of the final VLPs. To improve these issues, characterization techniques able to visualize and quantify the different steps of the process are needed. Fluorescence microscopy represents a powerful tool able to image multiple protein targets; however, its limited resolution hinders the study of viral constructs. Here, we propose the use of super-resolution microscopy and in particular of DNA-point accumulation for imaging in nanoscale topography (DNA-PAINT) microscopy as a characterization method for recombinant viral proteins on both cells and VLPs. We were able to quantify the amount of the three main influenza proteins (hemagglutinin (HA), neuraminidase (NA), and ion channel matrix protein 2 (M2)) per cell and per VLP with nanometer resolution and single-molecule sensitivity, proving that DNA-PAINT is a powerful technique to characterize recombinant viral constructs. ",

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note = "Funding Information: The authors would like to acknowledge the groups of Dr. Nico Callewaert and Dr. Xavier Saelens from VIB-UGENT Center for Medical Biotechnology for providing the influenza plasmids used in this paper (PHW2000 HA, NA, and M). M.A.-R. is grateful to the Spanish Ministry of Universities and the European Social Fund for the funding of his research activity by the research project (FPI16-SEV-2014-0425-16-1) (Ayudas para contratos predoctorales para la formaci{\'o}n de doctores 2016). S.P. and L.A acknowledge the financial support by the Spanish Ministry of Science and Innovation (PID2019-109450RB-I00/AEI/10.13039/501100011033), European Research Council/Horizon 2020 (ERC-StG-757397), “la Caixa” Foundation (ID 100010434), and the Generalitat de Catalunya (through the CERCA program and 2017 SGR 01536). ",

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Nanoscale Mapping of Recombinant Viral Proteins: From Cells to Virus-Like Particles

Abstract

Bibliographical note

Keywords

UN SDGs

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