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Updated: Jul 23, 2025

Avidity-based Extracellular Interaction Screening AVEXIS for the Scalable Detection of Low-affinity Extracellular Receptor-Ligand Interactions
Published on: March 5, 2012
An Interactome Assay for Detecting Interactions between Extracellular Domains of Receptor Kinases
Jente Stouthamer1, Sergio Martin-Ramirez1, Elwira Smakowska-Luzan2
1Wageningen University & Research Laboratory of Biochemistry, Wageningen, The Netherlands.
Abstract:
Interactions between extracellular domains (ECDs) are crucial for many physiological processes in the cell, most importantly perception of its environment. However, studying these often-transient interactions can be challenging. Here we describe a method that allows for in vitro detection of extracellular domain interactions through an oligomerization-based cell surface interaction (CSI) assay. In a CSI, bait- and prey-tagged proteins are produced and secreted by Drosophila S2 cells to ensure proper folding and post-translational modifications. Subsequently, Bait (FC fragment) and Prey (pentamer domain and alkaline phosphatase) tags allow the detection of interactions in protein A-coated 96 wells plates through a colorimetric readout. Due to the easy detection of interactions this approach can be used for high-throughput screening and mapping of extracellular interaction networks.
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