Investigation of biological cell-protein interactions using SPR sensor through laser scanning confocal...

Area of Science:

• Biophysics
• Cell Biology
• Immunology

Background:

• Investigating biological cell-protein interactions is crucial for understanding cellular processes and disease mechanisms.
• Existing methods may lack the sensitivity or real-time capabilities needed for dynamic interaction studies.
• Surface Plasmon Resonance (SPR) is a label-free technique sensitive to binding events at sensor surfaces.

Purpose of the Study:

• To develop and validate a novel integrated system combining laser scanning confocal imaging with surface plasmon resonance (LSCI-SPR) for studying cell-protein interactions.
• To demonstrate the capability of the LSCI-SPR system in real-time detection of antigen-antibody reactions involving suspended cells.
• To assess the potential applications of this LSCI-SPR method in cell biology and pharmacology.

Main Methods:

• A laser scanning confocal imaging-surface plasmon resonance (LSCI-SPR) system was developed.
• Mouse normal IgG was immobilized onto the SPR sensor chip surface.
• Suspension mouse lymphocyte cancer cells (L5178Y), labeled with Hoechst33342, were introduced to flow over the modified SPR chip.
• Fluorescence images and SPR signals were synchronously recorded in real-time as cell concentration varied.

Main Results:

• The LSCI-SPR system successfully recorded fluorescence images and SPR signals synchronously in real-time.
• An increase in the concentration of mouse lymphocyte cancer cells correlated with an increase in red fluorescence points.
• The observed changes in fluorescence intensity showed a strong correlation with the simultaneously recorded SPR signal.
• The method demonstrated the ability to detect cell-protein interactions via antigen-antibody binding on cell surfaces using different suspended cell types.

Conclusions:

• The developed LSCI-SPR system provides a novel and effective platform for investigating biological cell-protein interactions in real-time.
• The synchronous acquisition of imaging and SPR data allows for precise monitoring of binding events.
• This technique holds significant potential for applications in cell biology, drug discovery, and pharmacological studies involving cell surface interactions.