2.3 Protein microarray production

Sequence-verified, full-length cDNA expression plasmids in the T7-based mammalian expression vector pANT7_cGST were obtained from Arizona State University’s Biodesign Institute Plasmid Repository and are publicly available through DNASU (http://dnasu.asu.edu/DNASU/). The high-throughput preparation of high-quality supercoiled DNA for cell-free protein expression was performed as described [21]. Protein arrays displaying 10,247 human proteins were distributed evenly over five slides. Quality control was performed to determine inter and intra assay consistency by measuring DNA and protein expression (Figure 2 and Supplemental Table 1).

Quality control of protein microarrays. A. Quality control of DNA printing with picogreen. B. Quality control of in vitro protein production with anti-GST Mab staining. C. Reproducibility of protein signal intensity of two protein arrays produced in a single print batch.