A simple and high -performance immobilization technique of membrane protein from crude cell lysate sample for a membrane-based immunoassay application
Issued Date
2023-01-01
Resource Type
ISSN
15321819
eISSN
15324230
Scopus ID
2-s2.0-85141424094
Pubmed ID
36318041
Journal Title
Journal of Immunoassay and Immunochemistry
Volume
44
Issue
1
Start Page
76
End Page
89
Rights Holder(s)
SCOPUS
Bibliographic Citation
Journal of Immunoassay and Immunochemistry Vol.44 No.1 (2023) , 76-89
Suggested Citation
Khemthongcharoen N., Uawithya P., Yookong N., Chanasakulniyom M., Jeamsaksiri W., Sripumkhai W., Pattamang P., Juntasaro E., Kamnerdsook A., Houngkamhang N., Promptmas C. A simple and high -performance immobilization technique of membrane protein from crude cell lysate sample for a membrane-based immunoassay application. Journal of Immunoassay and Immunochemistry Vol.44 No.1 (2023) , 76-89. 89. doi:10.1080/15321819.2022.2137420 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/81912
Title
A simple and high -performance immobilization technique of membrane protein from crude cell lysate sample for a membrane-based immunoassay application
Other Contributor(s)
Abstract
Membrane 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.