Available Technologies

Background

Antibody probes used in immunological assays such as immunostaining and immunoassays generally bind strongly to antigens. Once such probes are bound, it is difficult to dissociate the antigen from the antibody probe. The inventors have established a multiplexed super-resolution imaging method called IRIS, which utilizes antibodies with low affinity but high specificity to the antigens. However, the acquisition of such antibody probes was challenging.

Description and Advantages

The inventors have shown that mutations introduced at conserved positions located at the base of the complementary determining region loops in the antibody probe can significantly reduce the half-life of antibody/antigen association while maintaining the binding specificity to the antigen.

• Automated acquisition of multiplexed super-resolution imaging is possible.

The antibody probe can be easily washed off with saline from the antigen (Fig. 1). Therefore, multiplexed super-resolution imaging is achieved by simply repeating the following three steps on the sample: Step 1: applying the antibody probe to the sample; Step 2: image acquisition; and Step 3: washing of the antibody probe (Fig. 2).

• Faithful special distribution of antigens can be captured

Because the antibody probe of the present invention dissociates from the antigen in a short period, steric hindrance due to the antibody probe bound to the antigen does not occur, unlike conventional antibody probes. This increases the labeling density of the antigen compared to the conventional method (PALM/STORM).