Structural and Kinetic Studies of Respiratory Syncytial Virus Neutralizing Antibodies
Engineering
BME 195
The project’s goal is to create atomic resolution models of the Respiratory Syncytial Virus (RSV) attachment protein bound to high affinity, neutralizing antibodies. These high resolution models will provide new and specific information about the molecular interactions between the antibody fragments and the attachment protein. These interactions can be used to determine which portions of the attachment protein are required to enhance viral infectivity. This work provides foundational research to develop a vaccine for this virus and other new therapeutics. A therapeutic to block attachment protein activity could decrease infection severity by 70 to 100-fold and provides a novel way to treat infection compared to current technology. Recombinant DNA cloning and expression were used to produce antibody fragments and short segments of the attachment protein in insect cells and E. coli. Affinity and Size Exclusion chromatography (SEC) were used to purify the antibody fragments and attachment protein segments. SEC was used to perform an initial screen of antibody fragment-attachment protein binding. Surface Plasmon Resonance (SPR) was used to quantify the binding interactions and compare to data from previous binding characterizations. SPR revealed binding affinity in the picomolar (10-12M) range for all three antibodies. Current work focuses on mutating attachment protein amino acids to better understand neutralizing and binding interactions between the binding antibodies and attachment proteins.
