Fabricating a UV-Vis and Raman Spectroscopy Immunoassay Platform

Area of Science:

• Biotechnology and Biomedical Engineering
• Analytical Chemistry
• Immunotechnology

Background:

• Immunoassays are vital tools in research and clinical diagnostics, utilizing antibody-antigen interactions for protein detection.
• Existing immunoassay infrastructure, including polystyrene plates and reagents for enzyme-linked immunosorbent assays (ELISA), is widely available.
• Standard ELISA kits have limitations in multiplexing capabilities due to spectral overlap in colorimetric and fluorescent detection methods.

Purpose of the Study:

• To address the limitations of current immunoassay techniques, particularly the low multiplexing capacity of ELISA.
• To provide a practical, do-it-yourself protocol for fabricating functionalized probes for Raman spectroscopy-based immunoassays.
• To enable researchers to develop custom multi-analyte detection assays by utilizing pre-established laboratory infrastructure.

Main Methods:

• Development of a protocol for preparing functionalized probes suitable for immunoassay applications.
• Integration of these probes with standard polystyrene plates for direct analyte detection.
• Utilizing both ultraviolet-visible (UV-Vis) spectroscopy and Raman spectroscopy for signal detection.

Main Results:

• The protocol facilitates the creation of probes for Raman spectroscopy-based immunoassays, offering superior multiplexing potential compared to traditional methods.
• Raman reporters exhibit enhanced photostability, reducing photobleaching issues common with fluorescent tags.
• The method allows for direct analyte detection using UV-Vis analysis and Raman spectroscopy in conjunction with standard plates.

Conclusions:

• This protocol empowers researchers to create their own advanced immunoassays, moving beyond the constraints of commercial kits.
• The approach enhances multi-analyte detection capabilities through the use of Raman spectroscopy, capitalizing on existing infrastructure.
• This DIY method offers a cost-effective and time-efficient solution for developing novel immunoassays for diverse research applications.