A simple and high -performance immobilization technique of membrane protein from crude cell lysate sample for a membrane-based immunoassay application

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dc.contributor.authorKhemthongcharoen, Numfon
dc.contributor.authorUawithya, Panapat
dc.contributor.authorYookong, Nutthapon
dc.contributor.authorChanasakulniyom, Mayuree
dc.contributor.authorJeamsaksiri, Wutthinan
dc.contributor.authorSripumkhai, Witsaroot
dc.contributor.authorPattamang, Pattaraluck
dc.contributor.authorJuntasaro, Ekachai
dc.contributor.authorKamnerdsook, Ampol
dc.contributor.authorHoungkamhang, Nongluck
dc.contributor.authorPromptmas, Chamras
dc.date.accessioned2026-08-06T10:40:38Z
dc.date.available2026-08-06T10:40:38Z
dc.date.issued2023-01-01
dc.description.abstractMembrane proteins are difficult to be extracted and to be coated on the substrate of the immunoassay reaction chamber because of their hydrophobicity. Traditional method to prepare membrane protein sample requires many steps of protein extraction and purification that may lead to protein structure deformation and protein dysfunction. This work proposes a simple technique to prepare and immobilize the membrane protein suspended in an unprocessed crude cell lysate sample. Membrane fractions in crude cell lysate were incorporated with the large unilamellar vesicle (LUV) that was mainly composed of POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine) before coating in the polystyrene plate by passive adsorption technique. Immunofluorescence staining and the Enzyme-Linked Immunosorbent Assay (ELISA) examination of a strictly conformation-dependent integral membrane protein, Myelin Oligodendrocyte Glycoprotein (MOG), demonstrate that LUV incorporated cell lysate sample obviously promotes MOG protein immobilization in the microplate well. With LUV incorporation, the dose–response curve of the MOG transfected cell lysate coating plate can be 2–9 times differentiated from that of the untransfected cell lysate coating plate. The LUV incorporated MOG transfected cell lysate can be efficiently coated in the microplate without carbonate/bicarbonate coating buffer assistance.
dc.identifier.citationJournal of Immunoassay and Immunochemistry, 44(1), 76-89, 2023
dc.identifier.doi10.1080/15321819.2022.2137420
dc.identifier.issn15321819
dc.identifier.other2-s2.0-85141424094
dc.identifier.urihttps://dspace.kmitl.ac.th/handle/123456789/14190
dc.sourceJournal of Immunoassay and Immunochemistry
dc.subjectconformation-dependent immunoassay
dc.subjectcrude cell lysate coating
dc.subjectenzyme-linked immunosorbent assay
dc.subjectMembrane protein immobilization
dc.subjectunilamellar vesicle preparation
dc.titleA simple and high -performance immobilization technique of membrane protein from crude cell lysate sample for a membrane-based immunoassay application
dc.typeArticle

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