In vitro exposure of proteins to ozone

Todd M. Umstead, David Phelps, Guirong Wang, Joanna Floros, Brian K. Tarkington

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

An in vitro system has been developed to expose proteins to ozone. The system is designed to deliver consistent and accurate levels of ozone over a range of concentrations (between 0.1 and ≥ 10 ppm) with extended exposure times (24 h or longer) in a humidified environment (100%). In the experiment presented in this article, ozone concentrations between 0.1 and 2.0 ppm were used. Ozone was generated by an electrical discharge ozonizer to ensure stability; it was continually monitored by an ultraviolet ozone analyzer and was precisely controlled by mass flow controllers, which gave reproducible results between runs. Humidity was closely regulated in the system to allow small amounts of protein solutions (50 μL or less) to be exposed without significant changes (<0.2%) in sample volume. The degree of surfactant protein-A (SP-A) oxidation by ozone was measured between runs to demonstrate the reproducibility of the system. A detailed description of the system is given, and protein oxidation detection methods and their limitations are discussed. Using these methods, we were able to assess oxidation of SP-A that apparently occurred prior to its isolation from the lung by bronchoalveolar lavage. This in vitro system allowed us to expose small amounts of protein to ozone in a simple, highly controlled, and reproducible manner.

Original languageEnglish (US)
Pages (from-to)1-16
Number of pages16
JournalToxicology Mechanisms and Methods
Volume12
Issue number1
DOIs
StatePublished - Jan 1 2002

Fingerprint

Ozone
Proteins
Pulmonary Surfactant-Associated Protein A
Bronchoalveolar Lavage
Oxidation
Humidity
In Vitro Techniques
Atmospheric humidity
Controllers

All Science Journal Classification (ASJC) codes

  • Toxicology
  • Health, Toxicology and Mutagenesis

Cite this

Umstead, Todd M. ; Phelps, David ; Wang, Guirong ; Floros, Joanna ; Tarkington, Brian K. / In vitro exposure of proteins to ozone. In: Toxicology Mechanisms and Methods. 2002 ; Vol. 12, No. 1. pp. 1-16.
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In vitro exposure of proteins to ozone. / Umstead, Todd M.; Phelps, David; Wang, Guirong; Floros, Joanna; Tarkington, Brian K.

In: Toxicology Mechanisms and Methods, Vol. 12, No. 1, 01.01.2002, p. 1-16.

Research output: Contribution to journalArticle

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