A modular method for rapidly prototyping targeted gas vesicle protein nanoparticles

Vassallo, R.; Ling, B.; Criado-Hidalgo, E.; Robinson, N.; Schrunk, E.; Liu, A.; Daghlian, G.; Li, H. R.; Swift, M. B.; Mannar, D.; Malounda, D.; Goldenberg, S. L.; Salcudean, S. E.; Shapiro, M. G.; Black, P. C.; Cox, M. E.

doi:10.1101/2025.08.06.668980

bioRxiv · 10.1101/2025.08.06.668980

A modular method for rapidly prototyping targeted gas vesicle protein nanoparticles

Vassallo, R.·Ling, B.·Criado-Hidalgo, E.·Robinson, N.·Schrunk, E.·Liu, A.·Daghlian, G.·Li, H. R.·Swift, M. B.·Mannar, D.·Malounda, D.·Goldenberg, S. L.·Salcudean, S. E.·Shapiro, M. G.·Black, P. C.·Cox, M. E.

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Abstract

Gas vesicles (GVs) are air-filled protein nanoparticles which are proving to be useful in a number of biomedical applications. We hypothesized that it could be possible to develop a modular method for creating rapidly prototyped GVs by modifying their surface chemistry to include targeting peptides in an orientation-specific manner. Here, we describe a modular method to create targeted GVs using His-tagged antibody fragments, ensuring that the antibody fragments are connected to the GV in an orientation-specific manner. This is achieved via the functionalization of the GVs with nickel-nitrilotriacetic acid (Ni-NTA) group. First, we validated that these functionalized GVs can bind His-tagged green fluorescent protein and characterized the particle size and surface charge of functionalized GVs. Then, GVs targeted to prostate-specific membrane antigen (PSMA) using a minibody were validated using a knockout validation in vitro.

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Vassallo, R., Ling, B., Criado-Hidalgo, E., Robinson, N., Schrunk, E., Liu, A., Daghlian, G., Li, H. R., Swift, M. B., Mannar, D., Malounda, D., Goldenberg, S. L., Salcudean, S. E., Shapiro, M. G., Black, P. C., Cox, M. E.. 2025-08-07. A modular method for rapidly prototyping targeted gas vesicle protein nanoparticles. https://doi.org/10.1101/2025.08.06.668980

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