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

Published: June 29, 2023

Abstract

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 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 (<str…

Representative Results

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 of aluminum foil (bottom layer, not shown). Blue dashed lines show the location of the cut lines. (C) Top and side view of …

Divulgazioni

The authors have nothing to disclose.

Materials

AcquireMP Refeyn MP data collection software
Anti-human thrombin Haematologic Technologies AHT-5020 RRID: AB_2864302
Cotton-tipped applicators Thorlabs CTA10 Cotton optical swabs for lens cleaning
Coverslips 24×24 mm Globe Scientific 1405-10
Coverslips 24×50 mm Fisher Scientific 12-544-EP
DiscoverMP Refeyn MP data processing software
Forceps Electron Microscopy Sciences 78080-CF Soft-tipped forceps for coverslips handling
Human α-thrombin Haematologic Technologies HCT-0020
Immersion oil Thorlabs MOIL-30
Isopropanol Alfa Aesar 36644
Microsoft Excel Microsoft Spreadsheet
OneMP Refeyn Mass Photometry instrument
Origin OriginLab Scientific graphing software
PBS Corning 46-013-CM 10x stock
Syringe filter Millipore SLGSR33SS Buffer and sample filtering

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Citazione di questo articolo
Mass Photometry to Study Antigen-Antibody Interactions on a Single-Molecule Level. J. Vis. Exp. (Pending Publication), e21393, doi: (2023).

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