Major
Biology
Anticipated Graduation Year
2025
Access Type
Restricted Access
Abstract
Binding affinity and binding specificity are two separate dimensions of molecular recognition. The former describes the affinity for the cognate ligand, while the latter describes the relative affinity of cognate to noncognate ligands. While the experimental determination of affinity is straightforward, it is much more difficult to measure specificity, since it requires the characterization of “off-target” binding to any of a large number of potential noncognate ligands. The goal of my research project is to use a new approach, immunoprecipitation-liquid chromatography mass spectrometry (IP-LC/MS), to characterize specificity of antibodies for their respective antigen as a function of their affinity maturation. This project may provide experimental evidence that the selection pressures during antibody affinity maturation act to increase not just affinity but also specificity for antigen, an important question in basic immunological research. Furthermore, it may help to better understand whether a subset of the mutations that occur during affinity maturation acts separately on affinity and specificity, or whether mutations more commonly affect both aspects of molecular recognition simultaneously. This knowledge might inform antibody engineering approaches that may need to optimize either for a given antibody to improve function for a given application, e.g. in antibody-based bioanalytical assays.
Faculty Mentors & Instructors
Joerg Zimmermann, Professor, Department of Chemistry and Biochemistry
Creative Commons License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 3.0 License.
Using Immunoprecipitation-mass spectrometry to characterize changes in specificity during antibody affinity maturation
Binding affinity and binding specificity are two separate dimensions of molecular recognition. The former describes the affinity for the cognate ligand, while the latter describes the relative affinity of cognate to noncognate ligands. While the experimental determination of affinity is straightforward, it is much more difficult to measure specificity, since it requires the characterization of “off-target” binding to any of a large number of potential noncognate ligands. The goal of my research project is to use a new approach, immunoprecipitation-liquid chromatography mass spectrometry (IP-LC/MS), to characterize specificity of antibodies for their respective antigen as a function of their affinity maturation. This project may provide experimental evidence that the selection pressures during antibody affinity maturation act to increase not just affinity but also specificity for antigen, an important question in basic immunological research. Furthermore, it may help to better understand whether a subset of the mutations that occur during affinity maturation acts separately on affinity and specificity, or whether mutations more commonly affect both aspects of molecular recognition simultaneously. This knowledge might inform antibody engineering approaches that may need to optimize either for a given antibody to improve function for a given application, e.g. in antibody-based bioanalytical assays.