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Capillary Electrophoresis to Monitor Peptide Grafting onto Chitosan Films in Real Time
Published on: October 26, 2016
An enzyme-responsive hydrogel of ferrocene-grafted carboxymethyl chitosan as a soft electrochemical sensor for MMP-9
Jinze Wang1, Haiqi Zhang1, Hongtao Hu1
1MOE Key Laboratory of Macromolecular Synthesis and Functionalization, Department of Polymer Science and Engineering, Zhejiang University, Hangzhou 310027, China.
Abstract:
Matrix metalloproteinase 9 (MMP-9) plays an important role in wound healing. However, overexpression of MMP-9 leads to the degradation of the newly formed extracellular matrix, which delays wound healing, ultimately leading to chronic wounds. Therefore, timely monitoring of the MMP-9 activity using simple, cost-effective methods is important to prevent the formation of chronic wounds. In this work, ferrocene-modified MMP-9 cleavage peptide (Fc-MG) modified carboxymethyl chitosan hydrogels were prepared as electrochemical biosensors. In the presence of MMP-9, the peptide chain is sheared, and the electrochemically active ferrocene segment is released. Therefore, analyzing the electrochemical activity of hydrogels using differential pulse voltammetry (DPV) can be used to determine MMP-9 activity. The results showed that the DPV peaks were correlated with the MMP-9 concentration in phosphate-buffered saline (PBS, pH 7.4) and Dulbecco's modified Eagle's medium (DMEM). Specifically, the corresponding coefficient of determination (R2) were 0.918 and 0.993. The limit of detections were 73.08 ng/mL and 131.71 ng/mL, respectively. Compared with the enzyme-linked immunosorbent assay, the hydrogel biosensor determined the concentration of MMP-9 in solution with simpler steps. This study demonstrates a novel strategy based on Fc-MG-modified hydrogels to monitor MMP-9 activity in cell secretion samples and shows the potential application in chronic wounds.
Insights
This study developed a novel hydrogel biosensor to detect matrix metalloproteinase 9 (MMP-9) activity, crucial for monitoring chronic wound healing. The simple, cost-effective electrochemical method accurately quantifies MMP-9 levels in biological samples.
Area of Science:
- Biomaterials Science
- Electrochemistry
- Wound Healing Research
Background:
- Matrix metalloproteinase 9 (MMP-9) is vital in wound healing, but its overexpression can impede healing by degrading extracellular matrix, leading to chronic wounds.
- Effective monitoring of MMP-9 activity is essential for preventing chronic wound development.
- Current monitoring methods can be complex and costly, necessitating simpler, more accessible alternatives.
Purpose of the Study:
- To develop a novel, cost-effective electrochemical biosensor for monitoring MMP-9 activity.
- To utilize ferrocene-modified MMP-9 cleavage peptide (Fc-MG) integrated into carboxymethyl chitosan hydrogels for MMP-9 detection.
- To evaluate the biosensor's performance in quantifying MMP-9 activity in relevant biological media.
Main Methods:
- Fabrication of carboxymethyl chitosan hydrogels modified with ferrocene-conjugated MMP-9 cleavage peptide (Fc-MG).
- Utilizing differential pulse voltammetry (DPV) to measure electrochemical activity changes upon MMP-9-mediated peptide cleavage.
- Testing the biosensor's response to varying MMP-9 concentrations in phosphate-buffered saline (PBS) and Dulbecco's modified Eagle's medium (DMEM).
Main Results:
- The DPV peak currents showed a strong correlation with MMP-9 concentrations in both PBS (R² = 0.918) and DMEM (R² = 0.993).
- Achieved limits of detection for MMP-9 were 73.08 ng/mL in PBS and 131.71 ng/mL in DMEM.
- The hydrogel biosensor offered a simpler detection process compared to traditional enzyme-linked immunosorbent assays (ELISA).
Conclusions:
- Fc-MG-modified hydrogel biosensors provide a sensitive and straightforward method for assessing MMP-9 activity.
- This electrochemical approach holds significant potential for real-time monitoring of MMP-9 in cell secretions.
- The developed biosensor offers a promising tool for managing and preventing chronic wounds.
