Surface immobilized hydrogels as versatile reagent reservoirs for microarrays

Area of Science:

• Biomolecular Engineering
• Analytical Chemistry
• Immunotechnology

Background:

• Antibody-based microarrays require minimizing cross-reactivity for accurate results.
• Current methods for reagent storage can be suboptimal for maintaining antibody and reagent integrity.

Purpose of the Study:

• To develop a versatile, compartmentalized storage system for antibodies and soluble microarray reagents.
• To improve the accuracy and reliability of antibody-based microarray assays.

Main Methods:

• Fabrication of hydrogel plugs (4% N,N-dimethylacrylamide, 3% bisacrylamide) immobilized on slides.
• Demonstration of hydrogel desiccation and rehydration capabilities for reagent storage.
• Utilizing hydrogels to prevent cross-reactivity and enable accurate quantification in biomarker assays.

Main Results:

• Hydrogels allow facile transfer of intact, functional biomolecules due to large pore sizes.
• Reagents stored in dry hydrogel microenvironments maintained performance.
• Achieved limits of detection comparable to conventional methods for HIV p24 antigen.
• Successfully prevented cross-reactivity for Staphylococcus aureus enterotoxins A and B.
• Enabled accurate quantification of interleukin 10 using a microarray-integrated calibration curve.

Conclusions:

• The described hydrogel system offers a versatile solution for compartmentalized reagent storage in microarrays.
• This approach enhances assay accuracy by mitigating cross-reactivity and preserving reagent functionality.
• The method holds potential for improving the reliability and performance of various microarray-based diagnostic applications.