3 research outputs found
Biobased, Internally pH-Sensitive Materials: Immobilized Yellow Fluorescent Protein as an Optical Sensor for Spatiotemporal Mapping of pH Inside Porous Matrices
The
pH is fundamental to biological function and its measurement therefore
crucial across all biosciences. Unlike homogenous bulk solution, solids
often feature internal pH gradients due to partition effects and confined
biochemical reactions. Thus, a full spatiotemporal mapping for pH
characterization in solid materials with biological systems embedded
in them is essential. In here, therefore, a fully biocompatible methodology
for real-time optical sensing of pH within porous materials is presented.
A genetically encoded ratiometric pH sensor, the enhanced superfolder
yellow fluorescent protein (sYFP), is used to functionalize the internal
surface of different materials, including natural and synthetic organic
polymers as well as silica frameworks. By using controlled, tailor-made
immobilization, sYFP is homogenously distributed within these materials
and so enables, via self-referenced imaging analysis, pH measurements
in high accuracy and with useful spatiotemporal resolution. Evolution
of internal pH is monitored in consequence of a proton-releasing enzymatic
reaction, the hydrolysis of penicillin by a penicillin acylase, taking
place in solution or confined to the solid surface of the porous matrix.
Unlike optochemical pH sensors, which often interfere with biological
function, labeling with sYFP enables pH sensing without altering the
immobilized enzyme’s properties in any of the materials used.
Fast response of sYFP to pH change permits evaluation of biochemical
kinetics within the solid materials. Thus, pH sensing based on immobilized
sYFP represents a broadly applicable technique to the study of biology
confined to the internally heterogeneous environment of solid matrices
Biobased, Internally pH-Sensitive Materials: Immobilized Yellow Fluorescent Protein as an Optical Sensor for Spatiotemporal Mapping of pH Inside Porous Matrices
The
pH is fundamental to biological function and its measurement therefore
crucial across all biosciences. Unlike homogenous bulk solution, solids
often feature internal pH gradients due to partition effects and confined
biochemical reactions. Thus, a full spatiotemporal mapping for pH
characterization in solid materials with biological systems embedded
in them is essential. In here, therefore, a fully biocompatible methodology
for real-time optical sensing of pH within porous materials is presented.
A genetically encoded ratiometric pH sensor, the enhanced superfolder
yellow fluorescent protein (sYFP), is used to functionalize the internal
surface of different materials, including natural and synthetic organic
polymers as well as silica frameworks. By using controlled, tailor-made
immobilization, sYFP is homogenously distributed within these materials
and so enables, via self-referenced imaging analysis, pH measurements
in high accuracy and with useful spatiotemporal resolution. Evolution
of internal pH is monitored in consequence of a proton-releasing enzymatic
reaction, the hydrolysis of penicillin by a penicillin acylase, taking
place in solution or confined to the solid surface of the porous matrix.
Unlike optochemical pH sensors, which often interfere with biological
function, labeling with sYFP enables pH sensing without altering the
immobilized enzyme’s properties in any of the materials used.
Fast response of sYFP to pH change permits evaluation of biochemical
kinetics within the solid materials. Thus, pH sensing based on immobilized
sYFP represents a broadly applicable technique to the study of biology
confined to the internally heterogeneous environment of solid matrices
Biobased, Internally pH-Sensitive Materials: Immobilized Yellow Fluorescent Protein as an Optical Sensor for Spatiotemporal Mapping of pH Inside Porous Matrices
The
pH is fundamental to biological function and its measurement therefore
crucial across all biosciences. Unlike homogenous bulk solution, solids
often feature internal pH gradients due to partition effects and confined
biochemical reactions. Thus, a full spatiotemporal mapping for pH
characterization in solid materials with biological systems embedded
in them is essential. In here, therefore, a fully biocompatible methodology
for real-time optical sensing of pH within porous materials is presented.
A genetically encoded ratiometric pH sensor, the enhanced superfolder
yellow fluorescent protein (sYFP), is used to functionalize the internal
surface of different materials, including natural and synthetic organic
polymers as well as silica frameworks. By using controlled, tailor-made
immobilization, sYFP is homogenously distributed within these materials
and so enables, via self-referenced imaging analysis, pH measurements
in high accuracy and with useful spatiotemporal resolution. Evolution
of internal pH is monitored in consequence of a proton-releasing enzymatic
reaction, the hydrolysis of penicillin by a penicillin acylase, taking
place in solution or confined to the solid surface of the porous matrix.
Unlike optochemical pH sensors, which often interfere with biological
function, labeling with sYFP enables pH sensing without altering the
immobilized enzyme’s properties in any of the materials used.
Fast response of sYFP to pH change permits evaluation of biochemical
kinetics within the solid materials. Thus, pH sensing based on immobilized
sYFP represents a broadly applicable technique to the study of biology
confined to the internally heterogeneous environment of solid matrices