Cryptophane Nanoscale Assemblies Expand 129 Xe NMR Biosensing

Analytical Chemistry
Serge D ZemerovIvan J Dmochowski

Abstract

Cryptophane-based biosensors are promising agents for the ultrasensitive detection of biomedically relevant targets via 129Xe NMR. Dynamic light scattering revealed that cryptophanes form water-soluble aggregates tens to hundreds of nanometers in size. Acridine orange fluorescence quenching assays allowed quantitation of the aggregation state, with critical concentrations ranging from 200 nM to 600 nM, depending on the cryptophane species in solution. The addition of excess carbonic anhydrase (CA) protein target to a benzenesulfonamide-functionalized cryptophane biosensor (C8B) led to C8B disaggregation and produced the expected 1:1 C8B-CA complex. C8B showed higher affinity at 298 K for the cytoplasmic isozyme CAII than the extracellular CAXII isozyme, which is a biomarker of cancer. Using hyper-CEST NMR, we explored the role of stoichiometry in detecting these two isozymes. Under CA-saturating conditions, we observed that isozyme CAII produces a larger 129Xe NMR chemical shift change (δ = 5.9 ppm, relative to free biosensor) than CAXII (δ = 2.7 ppm), which indicates the strong potential for isozyme-specific detection. However, stoichiometry-dependent chemical shift data indicated that biosensor disaggregation contributes to t...Continue Reading

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Citations

Aug 7, 2018·Chemphyschem : a European Journal of Chemical Physics and Physical Chemistry·Matthias SchnurrLeif Schröder
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Nov 16, 2018·Bioconjugate Chemistry·Jabadurai Jayapaul, Leif Schröder
Dec 29, 2020·Analytical Chemistry·Serge D ZemerovIvan J Dmochowski

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