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 first demonstrate that multiplexed proteins are displayed on M-NAPPA using protein-specific antibodies. Second, we compare the ability of M-NAPPA with non-multiplexed NAPPA to detect different protein-protein interactions and the serological antibody reactivity against 646 viral proteins. Next, we show the feasibility of M-NAPPA in performing high throughput screening for immune-dominant tuberculosis (TB) antigens through the use of an ultra-high density M-NAPPA TB proteome array containing four subarrays with 4,045 TB open reading frames (ORFs) on one slide. Using M-NAPPA TB protein microarrays, four new immune-dominant antigens in the sera of BCG-vaccinated guinea pigs were identified, which were then validated using ELISA. Finally, we propose a high throughput target discovery and verification pipeline based on the M-NAPPA approach.