Multiplatform Protein Detection and Quantification Using Glutaraldehyde-Induced Fluorescence for 3D Systems.

Area of Science:

• Biochemistry
• Materials Science
• Analytical Chemistry

Background:

• Glutaraldehyde (GTA) is a common biological fixative.
• GTA is known to induce fluorescence upon interaction with proteins, specifically bovine serum albumin (BSA).
• Existing protein quantification methods may lack sensitivity or cost-effectiveness for certain applications.

Purpose of the Study:

• To develop a novel, sensitive, and reproducible method for quantifying BSA.
• To utilize GTA-induced fluorescence for protein analysis.
• To establish a cost-effective and straightforward approach for BSA quantification, particularly within hydrogel systems.

Main Methods:

• Development of a BSA quantification method based on GTA crosslinking to induce fluorescence.
• Application of confocal laser scanning microscopy (CLSM), spectrofluorometry for quantitative assessment.
• Utilisation of widefield fluorescence microscopy for qualitative assessment.
• Preparation of alginate-methacrylate hydrogels containing varying BSA concentrations via photopolymerization.
• Incubation of hydrogels in glutaraldehyde solutions for fluorescence induction.

Main Results:

• Demonstrated a qualitative and quantitative correlation between BSA content and GTA-induced fluorescence.
• Achieved detection of BSA concentrations as low as 62.5 μg/mL using CLSM.
• Validated the method's sensitivity and reproducibility for protein quantification.

Conclusions:

• The developed GTA-induced fluorescence method offers a sensitive, reproducible, and cost-effective approach for BSA quantification.
• This technique is particularly advantageous for analyzing protein loading and release in three-dimensional hydrogel systems.
• The method is compatible with standard laboratory equipment, making it broadly accessible.