A bead-based multiplex immunoassay for cancer autoantibody biomarker discovery

• Published in: The Journal of immunology (1950) 2018-05, Vol.200 (1_Supplement), p.120-120.43
• Main Authors: Pepin, D, Godeny, Michael, Lie, Wen-Rong
• Format: Article
• Language: English
• ISSN: 0022-1767; 1550-6606
• DOI: 10.4049/jimmunol.200.Supp.120.43

Accurate, reliable and affordable blood tests are needed for early detection of cancer. Here we show data for the development and validation of a sensitive Luminex® bead-based multiplex immunoassay for detecting cancer autoantibodies in multiple cancer types. The cancer autoantibody biomarkers in this new multiplex panel include: CTAG1B/NY-ESO-1, Cyclin B/CCNB1, ENO1, Galectin-1, Galectin-3, Her2, HIF1α, HSP60, IMP2/IGF2BP2/p62, IMP3/IGF2BP3/KOC, MUC1, p16-INK4a, p53, SOX2, and Survivin/BIRC5. This multiplex immunoassay is sensitive and robust. In a 96-well plate, a mixture of recombinant protein-immobilized magnetic bead sets was incubated with serum samples for 2 hours with gentle shaking; after washing, the beads were incubated with detection antibody phycoerythrin (PE)-labeled anti-human IgG for 1.5 hours. After the final wash, the beads were resuspended in sheath fluid and the median fluorescent intensity (MFI) data of 50 beads per bead set were analyzed on a Luminex® reader. The performance of this panel was demonstrated using a set of serum samples from 78 cancer cases and 18 healthy controls. Elevated cancer autoantibodies were detected in serum from patients with various cancer types, as compared to healthy controls. In conclusion, this cancer autoantibody multiplex immunoassay has potential application as a non-invasive tool for studying early detection, diagnosis or monitoring of cancer.