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. 2016 Jul 14;11(7):e0159138. doi: 10.1371/journal.pone.0159138

Fig 2. Evaluation of the antibody quality in terms of spotting concentration, on-chip functionality and specificity.

Fig 2

(A) Two different scFvs antibodies, denoted as scFv1 (against complement protein C5) and scFv2 (against Myomesin 2), were dispensed in quintuplicate at three different concentrations, stock solution, 1:2 dilution and 1:4 dilution. The microarray was processed using a biotinylated serum sample. A scanner setting of 10 μm resolution, using 70% PMT gain and 90% laser power was used. (B) ScFv3 (anti-C1q) was spotted in 6 different concentrations, each in 12 replicate spots. The microarray was processed using pure antigen (5 nM pure Alexa 647-labeled C1q) and scanned at 10 μm resolution, using 60% PMT gain and 90% laser power. (C) Protein expression profiling of biotinylated serum sample. Several antibody clones directed towards the same antigen but against different epitopes were used, targeting Apolipoprotein-A4 (Apo-A4, n = 3), Complement factor 3 (C3, n = 6), Cystatin C (n = 4) and Monocyte Chemoattractant Protein-1 (MCP-1, n = 9). The observed signal intensities are given, in terms of fold change, in diseased vs. healthy samples.