Figure 2

Printing conditions and scFv antibody microarrays reproducibility. A) Printing map of a microarray comprising 384 scFvs against GFP or Trx. Controls used in the assay were: red box, mAb anti-T7Tag, 1:10 diluted. Yellow box, mAb anti-T7Tag 1:100 diluted. Blue box, crude Trx (top) or GFP (bottom) RTS extract 1:10 diluted. Grey box, printing buffer. Green box, TA4 anti-gastrin17 scFv; from right to left 1:10, 1:100, 1:1000 and 1:1000 dilutions. B) A representative image of a microarray probed with an anti-c-myc antibody to assess the correct printing of the scFvs. For detecting c-myc antibody, slides were incubated with Alexa Fluor 555-labeled goat anti-mouse IgG antibodies. White spots indicate a saturation of the green signal intensity. C) scFvs were spotted in duplicate onto FAST nitrocellulose coated slides to verify the intra-assay reproducibility. Replicated spots showed a uniform intensity either visually or by GenePix analysis. The two intensity values for each clone were quantified and plotted to assess the intra-array reproducibility.