REVisiting Antibody structures and repertoires through advances in Mass spectrometry and Proteomics

Introduction

In REVAMP, I will create innovative structural mass spectrometry (MS) and proteomics techniques to generate an unprecedented molecular understanding of the diversity and structural and functional characteristics of human antibody repertoires. These repertoires are highly personal and play a key role in our human immune response.

Research Focus

Recently, my research has taken a novel challenging avenue, with a focus on the development of cutting-edge MS-based techniques to better understand antibody structure and function. With these methods, I have started to monitor at the protein level changes in endogenous antibody repertoires with clonal resolution, including those induced by antigen-specific polyclonal responses in individuals.

Expansion of Techniques

Early studies were limited to IgG1, and in REVAMP, I aim to expand the toolbox to the full antibody repertoire, including all (sub)classes of antibodies. I aim to learn how and why antibody profiles in serum and other body fluids are alike or distinct, investigating:

1. Structural differences
2. Clonal differences
3. Functional differences over time and between individuals

Targeting Sub-Repertoires

I will develop technologies to target the sub-repertoires recognizing antigens from pathogens. Following identification by advanced top-down and bottom-up de novo sequencing, these antibodies will be followed up functionally as potential therapeutic leads.

Special Attention to IgM

Special attention will be given to IgM, the largest human antibody, appearing first in the response to an antigen. We recently made the preliminary striking observation that its structure is ill-defined in textbooks, and that human IgM may be quite different from IgM in other mammals.

Methodological Advances

This project will benefit from our recent advances in structural analysis by:

• Single molecule mass spectrometry
• Mass photometry
• Atomic force microscopy
• Electron microscopy

Conclusion

REVAMP will overhaul our current view of the structural diversity of our human antibody repertoire and thereby elevate our understanding of how the human immune system functions.