Mass Photometry to Study Antigen-Antibody Interactions on a Single-Molecule Level

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Samenvatting

Source: Wu, D. et. al., Rapid Determination of Antibody-Antigen Affinity by Mass Photometry. J. Vis. Exp. (2021)

In this video, we demonstrate the use of mass photometry to study antigen-antibody interactions in a flow chamber. The focused light scatters differently based on the mass of the molecules. A detector collects the scattered light and generates the interference pattern as spots of varied intensities, which correlate with the molecular masses.

Protocol

1. Prepare the flow chambers

Clean the glass coverslips
1. Using wash bottles with distilled water, ethanol, and isopropanol, rinse the 24 mm x 50 mm coverslips in the following order: water, ethanol, water, isopropanol, water. Dry the coverslips with a stream of clean nitrogen. It is important to rinse the coverslips from top to bottom, holding the bottom corner with soft-tipped forceps. Dry the coverslip in the same direction to avoid transferring contamination from the forceps (Figure 1A).
2. Similarly, rinse the 24 mm x 24 mm coverslips with distilled water, ethanol, and distilled water. Dry the coverslips with a strea....

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Resultaten

Coverslip preparation, double-sided tape usage diagram for microscopy experiments, fluid flow setup.
Figure 1: MP flow chamber preparation and loading. (A) Coverslip holding position for the cleaning procedure. (B) Alignment of the 24 x 24 mm coverslip (middle layer) and the double-sided tape (top layer) on the surface.......