Titration analysis of L514S-, L552S-, and NY-ESO-1-specific Abs in pleural effusions and sera of NSCLC patients. Patients’ pleural effusions (A) were diluted 50- to 31,250-fold, and sera (B) were diluted 250- to 781,250-fold with PBS containing 5% goat serum and 5% non-fat dry milk (NFDM). The samples were incubated in 96-well plates that had been protein-coated and blocked with 10% NFDM/PBS. After washing with 0.05% Tween/PBS, the plates were developed by HRP-conjugated antihuman IgG and substrate.