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Purification of an antitrypanosomal factor fromPseudomonas fluorescens by high-pressure liquid chromatography

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Abstract

Studies on the purification of an antitrypanosomal factor (ATF-II) obtained fromPseudomonas fluorescens disclosed that high-pressure liquid chromatography (HPLC) with a Rad-pak μporasil (10 μ silica) column and a GPC-60 Å column was an efficient procedure for the separation of the active components. Extraction of the factor with absolute ethanol prior to elution significantly enhanced the lytic activity of the HPLC eluates, as shown by marked pathologic changes followed by lysis in bioassays performed withTrypanosoma equiperdum. HPLC provided an increase of purification 30 times that obtained with gel filtration of the crude bacterial product. The lipopolysaccharide content of the purified fractions was markedly reduced and indicated an additional advantage for further in vivo tests in experimental infections withT. cruzi.

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Authors and Affiliations

  1. Laboratory of Parasitic Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland, USA

    Teresa I. Mercado

  2. Laboratory of Chemistry, National Institute of Diabetes, Digestive, and Kidney Diseases, National Institutes of Health, Bethesda, Maryland, USA

    Kenner C. Rice

  3. Pathology Branch, National Heart, Lung and Blood Institute, National Institutes of Health, Boilding 5, Room 210, 20892, Bethesda, Maryland, USA

    Victor J. Ferrans

  4. Waters Life Science Application Laboratory, Fairfax, Virginia, USA

    Patricia Strickler

Authors
  1. Teresa I. Mercado
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  2. Patricia Strickler
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  3. Kenner C. Rice
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  4. Victor J. Ferrans
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Mercado, T.I., Strickler, P., Rice, K.C. et al. Purification of an antitrypanosomal factor fromPseudomonas fluorescens by high-pressure liquid chromatography. Current Microbiology 16, 179–183 (1988). https://doi.org/10.1007/BF01568526

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  • DOI: https://doi.org/10.1007/BF01568526

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