Journal
CANCER NANOTECHNOLOGY
Volume 12, Issue 1, Pages -Publisher
SPRINGER WIEN
DOI: 10.1186/s12645-020-00071-7
Keywords
Pre-processing conditions; Gold nanoparticles; Protein corona; New molecular target proteins
Categories
Funding
- NIH [CA213278, CA136494]
- Peggy and Charles Stephenson Endowed Chair fund
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This study aimed to identify potential new molecular target proteins (NMTPs) responsible for poor outcomes in ovarian cancer. The results showed that pre-processing conditions can alter the protein corona (PC) composition and enhance the identification of NMTPs.
Background Ovarian cancer is one of the deadliest gynecological malignancies. While the overall survival of ovarian cancer patients has slightly improved in recent years in the developed world, it remains clinically challenging due to its frequent late diagnosis and the lack of reliable diagnostic and/or prognostic markers. The aim of this study was to identify potential new molecular target proteins (NMTPs) responsible for the poor outcomes. When nanoparticles (NP) are exposed to biological fluids, a protein coat, termed the protein corona (PC), forms around the NP, and the PC represents a tool to identify NMTPs. This study investigates the influence of pre-processing conditions, such as lysis conditions and serum/plasma treatment, on the PC composition and the resulting identification of NMTPs. Results Using gel electrophoresis, pre-processing conditions, including cell-lysis techniques and enrichment of low-abundance proteins (LAPs) by immunocentrifugation of serum/plasma, were shown to alter the relative amounts and compositions of proteins. PCs formed when 20 nm gold-NPs (GNPs) were incubated with lysate proteins from either RIPA- or urea lysis. Proteomic analysis of these PCs showed 2-22-fold enrichment of NMTPs in PCs from urea lysates as compared to RIPA lysates. Enriched NMTPs were then classified as cellular components, biological and molecular functions-associated proteins. The impact of enriched LAPs (eLAPs) on both PC composition and NMTP identification was shown by comparative proteomic analysis of original plasma, eLAPs, and PCs derived from eLAPs; eLAPs-PCs enhanced the abundance of NMTPs approximately 13%. Several NMTPs, including gasdermin-B, dermcidin, and kallistatin, were identified by this method demonstrating the potential use of this PC approach for molecular target discovery. Conclusion The current study showed that the pre-processing conditions modulate PC composition and can be used to enhance identification of NMTPs.
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