NULISA: a proteomic liquid biopsy platform with attomolar sensitivity and high multiplexing
Wei Feng, Joanne C. Beer, Qinyu Hao, Ishara S. Ariyapala, Aparna Sahajan, Andrei M. Komarov, Katie Cha, Mason Moua, Xiaolei Qiu, Xiaomei Xu, Shweta Iyengar, Thu Yoshimura, Rajini Nagaraj, Li Wang, Ming Yu, Kate Engel, Lucas Zhen, Wen Xue, Chen-jung Lee, Chan Ho Park, Cheng Peng, Kaiyuan Zhang, Adrian T. Grzybowski, Johnnie Hahm, Susanne V. Schmidt, Alexandru Odainic, Jasper Spitzer, Kasun Buddika, Dwight Kuo, Lei Fang, Bingqing Zhang, Steve Chen, Eicke Latz, Yiyuan Yin, Yuling Luo, Xiao-Jun Ma
Abstract
The blood proteome holds great promise for precision medicine but poses substantial challenges due to the low abundance of most plasma proteins and the vast dynamic range of the plasma proteome. Here we address these challenges with NUcleic acid Linked Immuno-Sandwich Assay (NULISA™), which improves the sensitivity of traditional proximity ligation assays by ~10,000-fold to attomolar level, by suppressing assay background via a dual capture and release mechanism built into oligonucleotide-conjugated antibodies. Highly multiplexed quantification of both low- and high-abundance proteins spanning a wide dynamic range is achieved by attenuating signals from abundant targets with unconjugated antibodies and next-generation sequencing of barcoded reporter DNA. A 200-plex NULISA containing 124 cytokines and chemokines and other proteins demonstrates superior sensitivity to a proximity extension assay in detecting biologically important low-abundance biomarkers in patients with autoimmune diseases and COVID-19. Fully automated NULISA makes broad and in-depth proteomic analysis easily accessible for research and diagnostic applications.