Home › Analytical Chem › Encapsulating Cytochrome c in Silica Aerogel Nanoarchitectures without Metal Nanoparticles while Retaining Gas-phase Bioactivity
Analytical Chem JoVE (Open Access) Citable · DOI

Encapsulating Cytochrome c in Silica Aerogel Nanoarchitectures without Metal Nanoparticles while Retaining Gas-phase Bioactivity

DOI: 10.3791/53802-v
What you'll learn
  • ✓ Encapsulate cytochrome c in silica sol-gels without metal nanoparticles
  • ✓ Process bioaerogels via supercritical drying to retain protein bioactivity
  • ✓ Detect nitric oxide gas-phase binding using spectroscopic analysis
Protocol

This procedure describes how to encapsulate cytochrome c (cyt. c) in silica (SiO2) sol-gels, process these gels to form bioaerogels, and use these bioaerogels to rapidly recognize nitric oxide (NO) through a gas-phase reaction. This type of protocol may aid in the future development of biosensors or other bioanalytical devices.

Difficulty
advanced
Total time
~3–5 days (including sol-gel synthesis, gelation, and supercritical drying cycles)
Biosafety
BSL-1

Steps

1
Prepare cytochrome c solutions and silica sol

Dissolve cytochrome c in appropriate buffer and synthesize silica sol through acid-catalyzed hydrolysis. Prepare gel molds for subsequent casting steps.

▶ 01:02
2
Mix cytochrome c with silica sol-gel precursor

Combine cytochrome c solution with silica sol under gentle agitation to form homogeneous cyt. c-silica sol-gels prior to gelation.

▶ 03:28
3
Supercritically dry cytochrome c-silica aerogels

Remove solvent from gelled samples using supercritical CO₂ drying to create porous aerogel architectures while preserving cytochrome c bioactivity.

▶ 06:10
4
Characterize aerogels and measure nitric oxide response

Perform spectroscopic analysis and gas-phase NO binding assays on final bioaerogel samples to confirm protein retention and biosensing capability.

▶ 09:46
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