An ultra-sensitive immunoassay for quantifying biomarkers in breast tumor tissue.
Urokinase-type plasminogen activator (uPA) and plasminogen activator inhibitor type-1 (PAI-1) have been validated at the highest level of evidence as clinical biomarkers of prognosis in breast cancer. The American Society of Clinical Oncology recommends using uPA and PAI-1 levels in breast tumors for deciding whether patients with newly diagnosed node-negative breast cancer can forgo adjuvant chemotherapy. The sole validated method for quantifying uPA and PAI-1 levels in breast tumor tissue is a colorimetric ELISA assay that takes 3 days to complete and requires 100-300 mg of fresh or frozen tissue. In this study we describe a new assay method for quantifying PAI-1 levels in human breast tumor tissue. This assay combines pressure-cycling technology to extract PAI-1 from breast tumor tissue with a highly sensitive liposome polymerase chain reaction immunoassay for quantification of PAI-1 in the tissue extract. The new PAI-1 assay method reduced the total assay time to one day and improved assay sensitivity and dynamic range by >100, compared to ELISA.
1. Laboratory of Proteomics and Protein Science, Baltimore Veterans Affairs Medical Center, Baltimore, MD, USA;
Carol B
CB
Fowler
2. Bon Secours Cancer Institute, Bon Secours Health System, Richmond, VA, USA.
Yan-Gao
YG
Man
1. Laboratory of Proteomics and Protein Science, Baltimore Veterans Affairs Medical Center, Baltimore, MD, USA;
Jeffrey T
JT
Mason
breast cancer; high-pressure tissue extraction; immunoassay; immunoliposome-PCR; plasminogen activator inhibitor type-1; tissue biomarkers