Ligand binding can affect the function of proteins and often causes conformational change in the protein target. Since form fits function, determining the secondary structure of proteins with and without ligands is essential for a more complete
understanding of how the ligand alters the protein’s structure and function. In this study the AQS3pro, developed by RedShiftBio, was used to measure protein secondary structure of 2 different ligands bound to their substrate protein.
The AQS3pro, featuring Microfluidic Modulation Spectroscopy (MMS), measures protein structure by combining infrared spectroscopy with microfluidics to enhance the sensitivity and accuracy of IR spectroscopy. Using a Quantum Cascade Laser (QCL)
that is 100 times brighter than FTIR light sources, MMS has enabled the ability to probe secondary structure more precisely and over a large concentration range (0.1-200+ mg/mL), all while using fully automated sample handling to minimize
instrument hands-on time and maximize sample throughput. Additionally, MMS uses a flow cell that modulates between sample and reference buffer, using real-time, automated buffer subtraction that is highly accurate and compatible for use with
complex buffer systems including organic modifiers such as DMSO.
MMS was used to investigate the stabilizing effect of two ligands on a protein substrate. In the data shown, the first ligand provides a strong stabilizing effect, whilst the second ligand provides a partial stabilization effect when the samples
have been exposed to higher temperatures. This data provides insights into the binding properties of the two ligands. The webinar demonstrates that overall, MMS is a sensitive secondary structure characterization tool that can enhance the
biophysical characterization toolbox by contributing secondary structural information to ligand binding applications.
Key Learning Objectives:
- The importance of determining the secondary structure of proteins with and without ligands for a more complete understanding of how the ligand alters the protein’s function
- How to probe secondary structure more sensitively at low concentrations and with buffers that contain DMSO
- How to detect structural changes caused by heat stress
- Pfizer evaluated The AQS3pro enabled by MMS technology to probe secondary structure more sensitively and over a wider concentration range than with FTIR. In this webinar, Alison H. Varghese, Principal Scientist, Structural and Molecular Sciences
part of Discovery Sciences, Pfizer, Inc will present the results and benefits for this study using Microfluidic Modulation Spectroscopy that demonstrates its ability to measure protein secondary structure more effectively than legacy techniques.
Alison H. Varghese
Principal Scientist, Structural and Molecular Sciences part of Discovery Sciences
Alison H. Varghese has a B.A. in biology from Rutgers University and an M.S. in cellular and molecular biology from Northeastern University. After a decade in academia, Alison joined Pfizer as a protein chemist. As Principal Scientist, she used
her protein chemistry and structural biology expertise to support medicinal design efforts in delivering effective medicines to patients in need. Alison is currently enrolled in Boston University’s graduate program in Project Management
and has co-authored over twenty peer-reviewed publications.
Valerie Ivancic is an Application Scientist at RedShiftBio. She was involved in one of the first Beta tests for the AQS3pro while in graduate school at Clark University in Worcester, MA. In graduate school, Valerie worked in a Biophysical Chemistry
lab gaining experience in circular dichroism, nuclear magnetic resonance spectroscopy, fluorescence spectroscopy, mass spectrometry, fast-performance liquid chromatography, transmission electron microscopy, and computer simulations in order
to address new ways of detecting, inhibiting, and degrading amyloid assemblies.
Cost: No Cost!