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Updated Method for In Vitro Analysis of Nanoparticle Hemolytic Properties

  1. Author:
    Neun, Barry
    Ilinskaya, Anna N
    Dobrovolskaia, Marina

  2. Author Address

    Cancer Research Technology Program, Nanotechnology Characterization Laboratory, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, P.O. Box B, Frederick, MD, 21702, USA., Cancer Research Technology Program, Nanotechnology Characterization Laboratory, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, P.O. Box B, Frederick, MD, 21702, USA. marina@mail.nih.gov.,
    1. Year: 2018
  2. Journal: Methods in molecular biology (Clifton, N.J.)
    1. 1682
    2. Pages: 91-102
  4. Type of Article: Article
  1. Abstract:

    Hemolysis is damage to red blood cells (RBCs), which results in the release of the iron-containing protein hemoglobin into plasma. An in vitro assay was developed and described earlier for the analysis of nanoparticle hemolytic properties. Herein, we present a revised version of the original protocol. In this protocol, analyte nanoparticles and controls are incubated in blood. Undamaged RBCs are removed by centrifugation and hemoglobin, released by the damaged erythrocytes, is converted to cyanmethemoglobin by incubation with Drabkin 39;s reagent. The amount of cyanmethemoglobin in the supernatant is measured by spectrophotometry. This measured absorbance is compared to a standard curve to determine the concentration of hemoglobin in the supernatant. The measured hemoglobin concentration is then compared to the total hemoglobin concentration to obtain the percentage of nanoparticle-induced hemolysis. The revision includes updated details about nanoparticle sample preparation, selection of nanoparticle concentration for the in vitro study, updated details about assay controls and case studies about nanoparticle interference with the in vitro hemolysis assay.

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  2. DOI: 10.1007/978-1-4939-7352-1_9
  3. PMID: 29039096

Library Notes

  1. Fiscal Year: FY2017-2018
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