sepharose and Pulmonary-Alveolar-Proteinosis

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.

Topics: Amniotic Fluid; Aspergillus fumigatus; Bronchoalveolar Lavage Fluid; Centrifugation; Chlorides; Chromatography, Affinity; Chromatography, Gel; Chromatography, Liquid; Edetic Acid; Electrophoresis, Polyacrylamide Gel; Glycoproteins; Humans; Leukocytes, Mononuclear; Manganese Compounds; Proteolipids; Pulmonary Alveolar Proteinosis; Pulmonary Surfactant-Associated Protein A; Pulmonary Surfactant-Associated Protein D; Pulmonary Surfactant-Associated Proteins; Pulmonary Surfactants; Sepharose; Urea