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Home / Equine Research Bank / Biochim Biophys Acta / Purification of horse eosinophil peroxidase.
Biochimica et biophysica acta1982; 701(2); 185-191; doi: 10.1016/0167-4838(82)90112-1

Purification of horse eosinophil peroxidase.

Abstract: Eosinophil peroxidase (donor: hydrogen-peroxidase oxidoreductase, EC 1.11.1.7) was isolated in a highly purified form (415/280 nm ratio, 1.05) from horse peripheral blood eosinophil. Eosinophil peroxidase was extracted from intact eosinophils (98-100% purity) or isolated eosinophil granules with 0.05 M acetate buffer (pH 4.7)/0.18 M NaCl and purified by chromatography on Sephadex G-200 and carboxymethylcellulose. Final elution was with 0.05 M acetate buffer (pH 4.7)/ 1 M NaCl. Horse eosinophil peroxidase is a strongly basic protein with bacterial properties when combined with H2O2 and iodide, bromide or, to a lesser degree, chloride.
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Publication Date: 1982-02-18 PubMed ID: 7041980DOI: 10.1016/0167-4838(82)90112-1Google Scholar: Lookup
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  • Journal Article
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  • Non-U.S. Gov't
  • Research Support
  • U.S. Gov't
  • P.H.S.

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research article discusses the process of successfully isolating and purifying the enzyme ‘eosinophil peroxidase’ from horse peripheral blood eosinophils, while also noting its basic properties and bacterial capabilities when combined with specified substances.

Research Objective

  • The main objective of this research was to purify the enzyme ‘eosinophil peroxidase’ from the eosinophils – a type of white blood cell in a horse’s peripheral blood. The desire for this purification process is largely due to the integral role that eosinophil peroxidase plays in the body’s immune response, tackling parasites and combating diseases.

Purification Method

  • Eosinophil peroxidase was extracted from either intact eosinophil (with 98-100% purity) or isolated eosinophil granules. This process was facilitated using a 0.05 M acetate buffer with a pH level of 4.7 and 0.18 M NaCl.
  • The extracted compound was then further purified through a process of chromatography. This involves the mixture being passed through a medium where the different components are separated due to their individual speeds of movement.
  • The researchers used two chromatography techniques – one on Sephadex G-200, and another on carboxymethylcellulose. Sephadex G-200 is a gel filtration media used in size exclusion chromatography, while Carboxymethylcellulose is a commonly used resin for ion-exchange chromatography.
  • Following this, the final elution of the purified eosinophil peroxidase was achieved using the same 0.05 M acetate buffer this time combined with 1 M NaCl.

Findings and Conclusion

  • The isolated horse eosinophil peroxidase was determined to be a strongly basic protein with certain bacterial indicators when combined with H2O2 – the chemical formula for hydrogen peroxide, and iodide, bromide, or to a smaller extent, chloride.
  • This observation suggested the potential use of horse eosinophil peroxidase in targeting bacterial infections, given its exhibited bacterial properties. However, further study would be required to explore this potential use case in more detail.

Cite This Article

APA
Jörg A, Pasquier JM, Klebanoff SJ. (1982). Purification of horse eosinophil peroxidase. Biochim Biophys Acta, 701(2), 185-191. https://doi.org/10.1016/0167-4838(82)90112-1

Publication

Biochimica et biophysica acta
ISSN: 0006-3002
NlmUniqueID: 0217513
Country: Netherlands
Language: English
Volume: 701
Issue: 2
Pages: 185-191

Researcher Affiliations

Jörg, A
    Pasquier, J M
      Klebanoff, S J

        MeSH Terms

        • Animals
        • Chromatography, Gel / methods
        • Chromatography, Ion Exchange
        • Cytoplasmic Granules / enzymology
        • Eosinophils / enzymology
        • Escherichia coli / drug effects
        • Horses / blood
        • Hydrogen-Ion Concentration
        • Peroxidases / blood
        • Peroxidases / isolation & purification
        • Peroxidases / pharmacology
        • Spectrum Analysis

        Citations

        This article has been cited 19 times.
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