Poster Presentation HUPO 2019 - 18th Human Proteome Organization World Congress

Bead-based workflows for rapid urinary proteome profiling and simultaneous recovery of DNA, RNA and proteins for clinical proteogenomic studies (#600)

Justin Jordaan 1 , Stoyan Stoychev 2 , Previn Naicker 2 , Ireshyn Govender 2 , Isak Gerber 2
  1. Resyn Biosciences, Edenvale, GAUTENG, South Africa
  2. NextGen Heath Cluster, CSIR, Pretoria, Gauteng, South Africa

The integration of cross-omics approaches, e.g. through genetic and protein isolation (proteogenomics), and analysis can provide comprehensive insights into the regulation of gene expression, cell signalling pathways, monitoring the onset of disease and subsequent progression. The collection and preservation of DNA, RNA and protein from the same patient sample is however challenging due to the different properties of these biomolecules and requirements for collection. This is often further complicated in clinical settings where minimal sample processing is possible due to a lack of suitable laboratory equipment. Urine is an attractive source for novel protein biomarker discovery due to its non-invasive collection and high abundance from patients. However, the DNA, RNA and protein content of urine is prone to degradation, and the dynamic range of e.g. proteins is greater than five orders of magnitude, with a wide molecular weight distribution due to protein cleavage that occurs in the kidneys. In this work we demonstrate workflows for the capture, processing and storage of DNA, RNA, and protein from patient samples that allow for robust, high-throughput sample processing. We compare two magnetic bead workflows for protein isolation, using hydrophilic affinity (HILIC) as well as strong anion exchange (SAX) for SPE, and benchmark against a traditional method and commercial kit. We further elaborate on the collection of proteins DNA and RNA from a range of biological samples, with downstream testing for a range of analyses.