Lumit: A Homogeneous Bioluminescent Immunoassay for Detecting Diverse Analytes and Intracellular Protein Targets

Traditional immunoassays to detect secreted or intracellular proteins can be tedious, require multiple washing steps, and are not easily adaptable to a high-throughput screening (HTS) format. To overcome these limitations, we developed Lumit, a novel immunoassay approach that combines bioluminescent...

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Published inMethods in molecular biology (Clifton, N.J.) Vol. 2612; p. 195
Main Authors Brian Hwang, Byounghoon, Alves, Juliano, Lazar, Dan, Nath, Nidhi, Engel, Laurie, O'Brien, Martha, Hsiao, Kevin, Kupcho, Kevin, Godat, Becky, Flemming, Rod, Goueli, Said, Zegzouti, Hicham
Format Journal Article
LanguageEnglish
Published United States 2023
Subjects
Cytokines
Humans
Immunoassay - methods
Immunologic Tests
Immunoassay
SARS-CoV2
Bioluminescence
Phosphorylation
NanoBiT
Antibody
Homogeneous
Cytokine
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Summary:Traditional immunoassays to detect secreted or intracellular proteins can be tedious, require multiple washing steps, and are not easily adaptable to a high-throughput screening (HTS) format. To overcome these limitations, we developed Lumit, a novel immunoassay approach that combines bioluminescent enzyme subunit complementation technology and immunodetection. This bioluminescent immunoassay does not require washes or liquid transfers and takes less than 2 h to complete in a homogeneous "Add and Read" format. In this chapter, we describe step-by-step protocols to create Lumit immunoassays for the detection of (1) secreted cytokines from cells, (2) phosphorylation levels of a specific signaling pathway node protein, and (3) a biochemical protein-protein interaction between a viral surface protein and its human receptor.
ISSN:1940-6029
DOI:10.1007/978-1-0716-2903-1_15