Colleen Ung,

Sponsor or Client: 

Title:

Developing a Highly-Multiplexed Serology Assay to Characterize the Mammalian Virosphere

Abstract:

Most emerging human infectious diseases are caused by pathogens originating from other animal hosts (i.e., zoonoses), and these pathogens have caused significant morbidity and mortality in humans. Anthropogenic activities (e.g., climate change, urbanization, deforestation) will continue to increase the risk of cross-species transmission as humans more frequently interact with animals and, by extension, the pathogens they carry. However, the diversity of viruses circulating in mammals remains poorly understood, impeding our ability to prepare for and mitigate future disease outbreaks. Because antibodies are often pathogen-specific and can persist long after infection, serological assays can be used to detect pathogen exposure months or years after it may have occurred. However, historically, serological assays have been limited to testing for antibody responses against one antigen at a time. With highly-multiplexed serology, antibody reactivity can be observed against 100,000s of antigens simultaneously. Our objective is to broadly apply highly-multiplexed serology to different mammal species to better understand the ecology of viruses that could spillover to humans. Here, we describe the design of an assay capable of broadly characterizing mammals’ antiviral antibody reactivity profiles, from which we can elucidate their history of viral exposures. Sequences were retrieved from NCBI and UniProt for our target viral families' focal proteins and clustered with a 70% identity threshold. For each cluster, 30 aa peptides were designed via a sliding window-set cover algorithm; oligonucleotide encodings were generated for the designed peptides. A 244,000 peptide library covering 27 viral families was created, with 2-16 focal proteins covered per family.