Abstract
A simple procedure has been developed for the purification of the surfactant proteins SP-A and SP-D from lung lavage of patients with alveolar proteinosis. The SP-D is purified by fractionation of the supernatant obtained after spinning the lavage at 10000 X g for 40 min, while the bulk of the SP-A is purified by fractionation of the pellet. The supernatant is applied to a maltosyl-agarose column and the bound SP-D is specifically eluted using MnCl2. The pellet is solubilised in 6 M urea and, following renaturation, the solubilised proteins are applied to maltosyl-agarose and SP-A eluted using a gradient of EDTA. Both SP-A and SP-D are further purified by gel-filtration on Superose-6. This procedure has also been used to prepare successfully human SP-A and SP-D from amniotic fluid and may be generally applicable to the isolation of these surfactant proteins from lung washings obtained from other species.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amniotic Fluid / chemistry*
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Aspergillus fumigatus
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Bronchoalveolar Lavage Fluid / chemistry*
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Centrifugation
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Chlorides
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Chromatography, Affinity
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Chromatography, Gel
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Chromatography, Liquid
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Edetic Acid
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Electrophoresis, Polyacrylamide Gel
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Glycoproteins / isolation & purification*
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Humans
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Leukocytes, Mononuclear / immunology
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Manganese Compounds
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Proteolipids / isolation & purification*
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Proteolipids / pharmacology
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Pulmonary Alveolar Proteinosis / metabolism*
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Pulmonary Surfactant-Associated Protein A
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Pulmonary Surfactant-Associated Protein D
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Pulmonary Surfactant-Associated Proteins
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Pulmonary Surfactants / isolation & purification*
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Pulmonary Surfactants / pharmacology
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Sepharose
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Urea
Substances
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Chlorides
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Glycoproteins
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Manganese Compounds
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Proteolipids
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Pulmonary Surfactant-Associated Protein A
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Pulmonary Surfactant-Associated Protein D
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Pulmonary Surfactant-Associated Proteins
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Pulmonary Surfactants
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Urea
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Sepharose
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Edetic Acid
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manganese chloride