An Ultra-Sensitive Immunoassay for Quantifying Biomarkers in Breast Tumor Tissue

作者：Fowler Carol B; Man Yan Gao; Mason Jeffrey T^*

来源：Journal of Cancer, 2014, 5(2): 115-124.

DOI：10.7150/jca.8084

• breast cancer
• plasminogen activator inhibitor type-1
• tissue biomarkers
• high-pressure tissue extraction
• immunoliposome-PCR
• immunoassay

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.

• 出版日期2014