A comparison of exosome purification methods using serum of Marek's disease virus (MDV)-vaccinated and -tumor-bearing chickens
Sabari Nath Neerukonda,
Nicholas A. Egan,
Joseph Patria,
Imane Assakhi,
Phaedra Tavlarides-Hontz,
Shannon Modla,
Eric R. Muñoz,
Matthew B. Hudson,
Mark S. Parcells
Affiliations
Sabari Nath Neerukonda
Department of Animal and Food Sciences, University of Delaware, Newark, DE 19716, USA
Nicholas A. Egan
Department of Biological Sciences, University of Delaware, Newark, DE 19716, USA
Joseph Patria
Department of Biological Sciences, University of Delaware, Newark, DE 19716, USA
Imane Assakhi
Department of Biological Sciences, University of Delaware, Newark, DE 19716, USA
Phaedra Tavlarides-Hontz
Department of Animal and Food Sciences, University of Delaware, Newark, DE 19716, USA
Shannon Modla
Delaware Biotechnology Institute, Bioimaging Center, Newark, DE 19711, USA
Eric R. Muñoz
Department of Kinesiology and Applied Physiology, University of Delaware, Newark, DE 19716, USA
Matthew B. Hudson
Department of Kinesiology and Applied Physiology, University of Delaware, Newark, DE 19716, USA
Mark S. Parcells
Department of Animal and Food Sciences, University of Delaware, Newark, DE 19716, USA; Department of Biological Sciences, University of Delaware, Newark, DE 19716, USA; Corresponding author.
Marek's disease (MD) is an alphaherpesvirus (Marek's disease virus, MDV)-induced pathology of chickens associated with paralysis, immunosuppression, neurological signs, and T-cell lymphomas. MD is controlled in poultry production via live attenuated vaccines. The purpose of the current study was to compare methods for precipitating exosomes from vaccinated and protected chicken sera (VEX) and tumor-bearing chicken sera (TEX) for biomarker analysis of vaccine-induced protection and MD lymphomas respectively. A standard polyethylene glycol (PEG, 8%) method was compared to a commercial reagent (total exosome isolation reagent, TEI) for exosome yield and RNA content. Although exosomes purified by PEG or TEI were comparable in size and morphology, TEI-reagent yielded 3-4-fold greater concentration. Relative expression of 8 out of 10 G. gallus- and MDV1-encoded miRNAs examined displayed significant difference depending upon the precipitation method used. Standard PEG yields comparable, albeit lower amounts of exosomes than the TEI-reagent and a distinctive miRNA composition.
