Advertisement

PEG Quantitation Using Reversed-Phase High-Performance Liquid Chromatography and Charged Aerosol Detection

  • Mackensie C. Smith
  • Jeffrey D. ClogstonEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1682)

Abstract

This chapter describes a method for the quantitation of polyethylene glycol (PEG) in PEGylated colloidal gold nanoparticles using a reversed-phase high-performance liquid chromatography (RP-HPLC) with charged aerosol detection. The method can be used to calculate the total PEG on the nanoparticle, as well as the bound and free unbound PEG fractions after a simple centrifugation step. This is a significant distinction as the bound PEG fraction affects biocompatibility, circulation time, and overall nanoparticle efficacy. PEG quantitation can be achieved through two methods, one involving the dissolution of colloidal gold nanoparticles by potassium cyanide (KCN) and the other by displacement of PEG by dithiothreitol (DTT). The methods outlined herein were applied to 30 nm colloidal gold grafted with 20 kDa PEG, but they can be easily adapted to any size colloidal gold nanoparticle and PEG chain length.

Key words

Polyethylene glycol (PEG) Surface characterization Charged aerosol detector Gold nanoparticles Stability Displacement Dissolution 

Notes

Acknowledgment

This project has been funded in whole or in part with Federal funds from the National Cancer Institute, National Institutes of Health, under Contract No. HHSN261200800001E. The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the U.S. Government.

References

  1. 1.
    Albanese A, Tang PS, Chan WCW (2012) The effect of nanoparticle size, shape, and surface chemistry on biological systems. Annu Rev Biomed Eng 14:1–16. doi: 10.1146/annurev-bioeng-071811-150124 CrossRefPubMedGoogle Scholar
  2. 2.
    Harris JM, Chess RB (2003) Effect of pegylation on pharmaceuticals. Nat Rev Drug Discov 2(3):214–221. doi: 10.1038/Nrd1033 CrossRefPubMedGoogle Scholar
  3. 3.
    Jokerst JV, Lobovkina T, Zare RN, Gambhir SS (2011) Nanoparticle PEGylation for imaging and therapy. Nanomedicine 6(4):715–728. doi: 10.2217/Nnm.11.19 CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Yowell SL, Blackwell S (2002) Novel effects with polyethylene glycol modified pharmaceuticals. Cancer Treat Rev 28:3–6. doi: 10.1016/S0305-7372(02)80002-0 CrossRefPubMedGoogle Scholar
  5. 5.
    Almeida JP, Figueroa ER, Drezek RA (2014) Gold nanoparticle mediated cancer immunotherapy. Nanomedicine 10(3):503–514. doi: 10.1016/j.nano.2013.09.011 CrossRefPubMedGoogle Scholar
  6. 6.
    Blanco E, Hsiao A, Mann AP, Landry MG, Meric-Bernstam F, Ferrari M (2011) Nanomedicine in cancer therapy: innovative trends and prospects. Cancer Sci 102(7):1247–1252. doi: 10.1111/j.1349-7006.2011.01941.x CrossRefPubMedGoogle Scholar
  7. 7.
    Cai W, Gao T, Hong H, Sun J (2008) Applications of gold nanoparticles in cancer nanotechnology. Nanotechnol Sci Appl 2008(1):17–32. doi: 10.2147/NSA.S3788 CrossRefGoogle Scholar
  8. 8.
    Jain S, Hirst DG, O'Sullivan JM (2012) Gold nanoparticles as novel agents for cancer therapy. Br J Radiol 85(1010):101–113. doi: 10.1259/bjr/59448833 CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    van Vlerken LE, Amiji MM (2006) Multi-functional polymeric nanoparticles for tumour-targeted drug delivery. Expert Opin Drug Deliv 3(2):205–216. doi: 10.1517/17425247.3.2.205 CrossRefPubMedGoogle Scholar
  10. 10.
    Smith MC, Crist RM, Clogston JD, McNeil SE (2015) Quantitative analysis of PEG-functionalized colloidal gold nanoparticles using charged aerosol detection. Anal Bioanal Chem 407(13):3705–3716. doi: 10.1007/s00216-015-8589-2 CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2018

Authors and Affiliations

  1. 1.Cancer Research Technology Program, Nanotechnology Characterization LaboratoryLeidos Biomedical Research, Inc., Frederick National Laboratory for Cancer ResearchFrederickUSA

Personalised recommendations