Diagnosis of oesophageal cancer currently requires expensive and invasive endoscopy-biopsy procedures, which are often only performed when obvious symptoms have manifested (usually at a late stage of the disease). QIMR Berghofer Medical Research Institute and Proteomics International have partnered to develop a targeted mass spectrometry-based oesophageal adenocarcinoma diagnostic test using serum samples. Panels of lectin-pulled down serum protein biomarkers have been found by the first laboratory that correlate with the presence of early stage oesophageal adenocarcinoma (high grade dysplasia). The second laboratory has automated and optimised the methodology to measure 33 target peptides in the lectin pulldown in a 20 minute microflow LCMS-MRM run. Analysis of an initial cohort (n=50) shows the method to be robust and reproducible with an average intraday CV of 9.3% across the 33 peptides and an average interday CV of 11.5%. A further larger cohort (n=266) has been analysed, and is being used to validate the initial data and build a statistical model to maximise the predictive power of the panel of protein biomarkers. The outcome of this collaborative effort is to produce a clinically viable diagnostic test to support screening and early detection in populations at high risk of oesophageal adenocarcinoma.