Multiplexed Nucleic Acid Programmable Protein Arrays

Yu, Xiaobo; Song, Lusheng; Petritis, Brianne; Bian, Xiaofang; Wang, Haoyu; Viloria, Jennifer; Park, Jin; Bui, Hoang; Li, Han; Wang, Jie; Liu, Lei; Yang, Liuhui; Duan, Hu; McMurray, David N.; Achkar, Jacqueline M.; Magee, Mitch; Qiu, Ji; LaBaer, Joshua

We purified 646 viral ORF plasmids from ~23 viruses, normalized their concentrations to 1,200 ng/µL, and printed viral NAPPA and M-NAPPA arrays in duplicate [20] . Analyses of the deposited DNA and displayed protein levels indicate that most viral DNA plasmids were successfully printed, expressed, and captured onto the microarrays in a reproducible manner (Figure 3A) . For example, plasmid DNA deposition across technical replicates of NAPPA and M-NAPPA had correlations (R) of 0.95 and 0.96, respectively. The protein display correlation (R) across technical replicates of NAPPA and M-NAPPA were 0.90 and 0.93, respectively ( Figure 3B, Figure S6 ). "Non-spots" containing printing buffer alone without plasmid DNA was used as a negative control. 94% and 93% of the spots on NAPPA and M-NAPPA viral arrays, respectively, produced signal that was at least two SDs above the average signal intensity of these "non-spots" (Figure S7 ). Together with Figure 2 , the results indicate that the majority of viral proteins can be displayed on M-NAPPA arrays.