Identification of Brain ECM Binding Variable Lymphocyte Receptors Using Yeast Surface Display.

Benjamin J Umlauf, John S Kuo, Eric V Shusta

Methods in molecular biology (Clifton, N.J.) 2022

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Extracellular matrix (ECM) is a rich mixture of proteins and glycans secreted by cells. This includes typical ECM structures such as collagen and heparin as well as glycosylated, secreted proteins such as growth factors and peptidases. Certain components of ECM are ubiquitous among all tissue; however, each biological tissue also displays unique variations that can be identified using biopanning techniques. Here we describe using a variable lymphocyte receptor (VLR) yeast surface display library to identify selective binders to brain ECM by combining ECM biopanning with a rapid ELISA-based screen using clonal VLRs isolated directly from the yeast surface. Finally, potential ECM-binding candidates can be verified by immunostaining murine brain sections with VLRs released from the yeast surface. These methods provide a framework for the identification of tissue-selective ECM-binding VLRs using yeast surface display techniques and could easily be adapted for other binding scaffolds or ECM from other tissues. © 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.

Citation

Benjamin J Umlauf, John S Kuo, Eric V Shusta. Identification of Brain ECM Binding Variable Lymphocyte Receptors Using Yeast Surface Display. Methods in molecular biology (Clifton, N.J.). 2022;2491:235-248