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Cellular and Molecular Bioengineering

Cellular and Molecular Bioengineering - Poster Session B

Poster I11 - Identification of Nanobodies for the Observation of Synapse Structure

Thursday, October 24, 2024

2:45 PM - 3:45 PM EST

Location: Exhibit Hall E, F & G

Presenting Author(s)

Nathan Irvin (he/him/his)

Undergraduate Student in Biomedical Engineering
West Virginia University
Morgantown, West Virginia, United States

Co-Author(s)

Candis Dancy (she/her/hers)

Graduate Student
West Virginia University
Morgantown, West Virginia, United States
KH

Kaitlyn Heintzelman

MD/PhD Student in Biomedical Engineering
West Virginia University, United States

Primary Investigator(s)

Moriah Katt, PhD (she/her/hers)

Assistant Professor
West Virginia University, United States

Introduction: The synapse is the junction between one neuron's axon terminal and another's dendrite where signals are transferred from one neuron to the next. The synapse modulates information transfer and storage within the brain by undergoing structural changes such as alterations in protein expression and localization. These changes play a crucial role in CNS disorders, yet little is known about them. Understanding how the synaptic structure changes in disease may enable the development of novel therapeutic approaches. Due to the density of the proteins in the synapse traditional IgG antibodies have been shown to be less effective due to their large size. Nanobodies (Nbs) offer a valuable potential alternative for synaptic labeling due to their relatively small size. Using yeast surface display we will screen a library of nanobodies for synaptic binding nanobodies for enhanced detection of synaptic structures.

Materials and

Methods: First, we used yeast surface display (YSD) to construct a naive nanobody library of size 1x10^8. The library was screened against D21 cortical rat neurons in 4 rounds of biopanning with enrichment of yeast binding clones in between each round. Lead clones were identified and tested for binding as individual clones. Plasmids containing the Nbs were extracted from yeast to determine the diversity of the library and to clone individual Nbs for further analysis as double proteins.

Results, Conclusions, and Discussions: After 4 rounds of biopanning we saw significant enrichment of the neuron-binding yeast. Of the 95 individual clones we were able to isolate, we have identified 22 high-binding, 29 medium-binding, and 44 low-binding clones for further analysis. These nanobodies will ultimately offer a tool for scientists to localize and study individual pre- and postsynaptic proteins.

Acknowledgements (Optional):