RPPA Reveals Altered Phosphorylation Profile in Plasmodium-infected Hepatocytes

IN ONCYTE nitrocellulose film slides

Introduction

Plasmodium parasites inflict widespread global disease, which in the case of malaria is caused by the intraerythrocytic form of the parasite.  The parasite infects the host by traveling to the liver where it intracellularly develops and affects hepatocyte cells, initiating a range of responses including autophagy and apoptosis.  Different hosts, and even different hepatocyte cells within a single host exhibit different capacities to support or limit infection, the cause of which is not comprehensively understood.  Kaushansky, et al, investigated differential susceptibility to infection of hepatocytes populations of BALB/cJ and BALB/cByJ mice, along with elevated ploidy level mice cells were evaluated to identify conserved factors that regulate infection using RPPA.  The RPPA assay was constructed using ONCYTE nitrocellulose slides arrayed with protein lysates of the different hepatocyte populations with Aushon contact pin printing methods.  These arrays were probed with a range of antibodies to quantitate total or modified protein abundance and normalized using beta-actin measurements.  A number of differentially expressed and upregulated proteins and post-translational modifications were identified using this high-throughput method most notably phospho-modified RPS6 (pS235/236, pS236/236) and SOD2.  Results were validated using orthogonal methods which also illustrates the accuracy of the RPPA method.

RPPA and ONCYTE® Nitrocellulose Film Slides

The RPPA assay platform is a powerful tool for protein expression profiling, biomarker discovery, pharmacodynamics evaluation, and cancer diagnostics. Porous nitrocellulose (PNC) film slides are the optimal protein microarray substrate technology which has a unique combination of engineered characteristics to provide a powerful high-throughput tool for economic and accurate lysate profiling:

  • Maximum binding capacity from optimized 3D pore structure
  • Optimal protein specificity and reactivity provided by nitrocellulose preservation of native protein configurations
  • Superior sensitivity produced by high signal to noise of fluorescent light scattering
  • Quantitative results from broad dynamic range of protein binding
  • Tremendous throughput provided by flexible configurations and high-density multiplex capability
  • Flexible compatibility with colorimetric, chemiluminescent, fluorescent and near-IR detection strategies
  • Reproducible results from high quality ISO 9001/13485 production processes

Learn more here:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6447308/