Serum from lung adenocarcinoma cancer patients treated in Shanghai Pulmonary Hospital were banked according to protocols approved by Institutional Review Board (K16–245-1). EGFR mutations were detected by ARMS method using ADx EGFR Mutations Detection Kit (Amoy Diagnostics, Xiamen, China) [40 (link)].
The recombinant EGFR protein (extracellular part aa 1–645 and intracellular part aa 668–1210) was from Sinobiologicals, China. The EGFR protein was bound to ELISA plates (1 μg/ml) for overnight at 4 °C. 100 μl serum of lung cancer patients were added and incubated for 1 h at room temperature. The plates were washed for 3 times by washing buffer (PBS with 0.05% Tween-20), and incubated with HRP labeled goat anti-human IgG for 1 h, followed by colorimetric detection. PBS 1% BSA was used as blank for determining the cutoff value.
The recombinant EGFR protein (extracellular part aa 1–645 and intracellular part aa 668–1210) was from Sinobiologicals, China. The EGFR protein was bound to ELISA plates (1 μg/ml) for overnight at 4 °C. 100 μl serum of lung cancer patients were added and incubated for 1 h at room temperature. The plates were washed for 3 times by washing buffer (PBS with 0.05% Tween-20), and incubated with HRP labeled goat anti-human IgG for 1 h, followed by colorimetric detection. PBS 1% BSA was used as blank for determining the cutoff value.
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