Nanoprojectile Secondary Ion Mass Spectrometry for Analysis of Extracellular Vesicles. Academic Article uri icon

abstract

  • We describe a technique based on secondary ion mass spectrometry with nanoprojectiles (NP-SIMS) for determining the protein content of extracellular vesicles, EVs, via tagged antibodies. The technique uses individual gold nanoprojectiles (e.g., Au4004+ and Au28008+), separated in time and space, to bombard a surface. For each projectile impact (10-20 nm in diameter), the co-emitted molecules are mass analyzed and recorded as an individual mass spectrum. Examining these individual mass spectra for co-localized species allows for nanoscale mass spectrometry to be performed. The high lateral resolution of this technique is well suited for analyzing nano-objects. SIMS is generally limited to analyzing small molecules (below ∼1500 Da); therefore, we evaluated three molecules (eosin, erythrosine, and BHHTEGST) as prospective mass spectrometry tags. We tested these on a model surface comprising a mixture of all three tags conjugated to antibodies and found that NP-SIMS could detect all three tags from a single projectile impact. Applying the method, we tagged two surface proteins common in urinary EVs, CD63 and CD81, with anti-CD63-erythrosine and anti-CD81-BHHTEGST. We found that NP-SIMS could determine the relative abundance of the two proteins and required only a few hundred or thousand EVs in the analysis region to detect the presence of the tagged antibodies.

published proceedings

  • Anal Chem

altmetric score

  • 1

author list (cited authors)

  • Verkhoturov, D. S., Crulhas, B. P., Eller, M. J., Han, Y. D., Verkhoturov, S. V., Bisrat, Y., Revzin, A., & Schweikert, E. A.

citation count

  • 1

complete list of authors

  • Verkhoturov, Dmitriy S||Crulhas, Bruno P||Eller, Michael J||Han, Yong D||Verkhoturov, Stanislav V||Bisrat, Yordanos||Revzin, Alexander||Schweikert, Emile A

publication date

  • May 2021