High throughput assay for detection of antibodies targeting multiple subunit membrane proteins of human neuron synapse in vitro

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Presentation Time: 03:15 PM - 04:30 PM
Poster Board Number: B894
Abstract ID: 4952

Presenting Author:
Quanzhen Li , Chief Technology Officer at GeneCopoeia, Univ. of Texas Southwestern Med. Ctr.

Abstract:

Synapses are essential to the transmission of nervous impulses from one neuron to another. There are more than 1000 multiple-subunit membrane proteins in synapse. Some of them are drug targets and/or autoantigens. Most autoantibodies against synapse membrane proteins recognize conformation of extracellular domains. Based on known structures and predicted structures by chatGPT, we used N-methyl-D-aspartate (NMDA)receptor as model for quantitative assay in vitro. In order to keep the native form of the extracellular domains of NMDA receptor, we modified some amino acids in transmembrane domains and intracellular domains, establishing a stable cell line expressing cluster NMDA receptor used to sensitively detect autoantibody in human serum and cerebrospinal fluid.  We also prepared stable NMDA receptor tetramer complex from the stable cell line and prepared monoclonal antibodies (mAB) against subunit GluN1 and GluN2. Based on tetramer complex and mABs, we established a method to quantitatively detect antibodies from clinical samples.