Development of a lateral flow assay for the detection of β-actin in whole cell extracts



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The primary objective of this summer project was to gain hands-on experience in Lateral Flow Assay (LFA) development for cancer diagnostics and more specifically the development of an LFA for the detection of β-actin in whole cell extracts. Initially, a model LFA for the detection of human Chorionic Gonadotropin (hCG) was used. Then, an LFA for the detection of β-actin in CA46 human B-cell lymphoma cells was developed. The Lateral Flow Assay is a rapid, point-of-care immunodiagnostic test commonly seen as the home pregnancy test. This method of diagnosis is convenient due its user-friendly format and ease of use without specialized personnel. In typical LFAs, gold nanoparticles are conjugated to antibodies that capture the target analyte from the sample. As the sample flows through the LFA membrane, the gold nanoparticle-antibody-target complex is captured onto the antibodies immobilized (test line) on the membrane of the strip. Detection of β-actin in whole cell extracts allows researchers to confirm reproducibility of sample preparation and normalization of the number of cells in specimens. Thus, a β-actin LFA can serve as a control in a rapid Acute Promyelocytic Leukemia (APL) LFA test for blood, which is under development in Professor Willson’s lab at the University of Houston.