Dual Staining for Sensitive Detection of Antigen-Specific B Cells

The Power of Dual Staining: High Sensitivity and Minimal Background

In B cell detection, non-specific binding and background is a known challenge resulting from the stickiness of naïve and activated B cells, among others. Dual staining is a way to increase reliability in your B cell detection. Klickmer® is now provided with BV421 in addition to FITC, PE, and APC, providing more options for dual staining of antigen-specific B cells.

Proof of Concept Study: Spike-specific B Cells After COVID-19 Vaccination

Klickmer® technology can easily be conjugated with mono-biotinylated molecules of your choice for tailored B cell detection. In this study, Klickmer® BV421 and PE reagents were assembled with SARS-CoV-2 spike proteins for longitudinal detection and phenotyping of antigen-specific B cells after vaccination.

The assembly protocol with spike proteins can be found in our “Application Protocol - Klickmer® for Detection of SARS-CoV-2 Spike-specific B cells” (see immudex.com/resources/protocols).

The study relied on PBMC samples from Immudex’s COVID-19 Biobank, designed to support longitudinal studies of vaccinated individuals.

Detecting Vaccination Responses Over Time

Using dual staining with spike-Klickmer® BV421 and PE, we could successfully detect spike-specific B cells and observe increased numbers with each vaccination with minimal impact of background. The largest increase was observed after the third dose highlighting a robust immune response to the spike protein of SARS-CoV-2, providing crucial insights into the efficacy of multiple vaccine doses.

Monitoring Phenotypic Changes in B Cells

Beyond just detecting B cell responses, in combination with an antibody panel, Klickmer® BV421 and PE enabled tracking of phenotypic shifts over time. Notably, many spike-specific B cells transitioned to an IgD-CD27+ phenotype, a hallmark of memory B cells, which are critical for long-term immunity. This shift underscores the lasting impact of the vaccination on the immune system, providing deeper insights into how the immune response evolves post-vaccination.

Conclusions

By leveraging the dual staining power of BV421 with Klickmer® Technology, we could achieve highly precise detection of antigen-specific B cells while minimizing background. Klickmer® technology can provide new and more reliable ways of understanding the dynamics of the immune system, effectiveness of vaccines and studying long-term development of memory B cell populations.

Related Resources

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BV421 is equivalent to Brilliant Violet™ 421, which is a trademark or registered trademark of Becton, Dickinson and Company or its affiliates, and is used under license. Powered by BD Innovation.