Efficient delipidation of a recombinant lung surfactant lipopeptide analogue by liquid-gel chromatography

Autoři: Oihana Basabe-Burgos aff001;  Jakub Zebialowicz Ahlström aff001;  Pavol Mikolka aff001;  Michael Landreh aff003;  Jan Johansson aff001;  Tore Curstedt aff004;  Anna Rising aff001
Působiště autorů: Department of Neurobiology, Care Sciences and Society, Division for Neurogeriatrics, Karolinska Institutet, Huddinge, Sweden aff001;  Biomedical Center Martin and Department of Physiology, Jessenius Faculty of Medicine in Martin, Comenius University in Bratislava, Martin, Slovakia aff002;  Science for Life Laboratory, Department of Microbiology, Tumour and Cell Biology, Karolinska Institutet, Tomtebodavägen, Stockholm, Sweden aff003;  Department of Molecular Medicine and Surgery, Karolinska Institutet at Karolinska University Hospital, Stockholm, Sweden aff004;  Department of Anatomy, Physiology and Biochemistry, Swedish University of Agricultural Sciences, Uppsala, Sweden aff005
Vyšlo v časopise: PLoS ONE 14(12)
Kategorie: Research Article
doi: https://doi.org/10.1371/journal.pone.0226072


Pulmonary surfactant preparations extracted from natural sources have been used to treat millions of newborn babies with respiratory distress syndrome (RDS) and can possibly also be used to treat other lung diseases. Due to costly production and limited supply of animal-derived surfactants, synthetic alternatives are attractive. The water insolubility and aggregation-prone nature of the proteins present in animal-derived surfactant preparations have complicated development of artificial surfactant. A non-aggregating analog of lung surfactant protein C, SP-C33Leu is used in synthetic surfactant and we recently described an efficient method to produce rSP-C33Leu in bacteria. Here rSP-C33Leu obtained by salt precipitation of bacterial extracts was purified by two-step liquid gel chromatography and analyzed using mass spectrometry and RP-HPLC, showing that it is void of modifications and adducts. Premature New Zealand White rabbit fetuses instilled with 200mg/kg of 2% of rSP-C33Leu in phospholipids and ventilated with a positive end expiratory pressure showed increased tidal volumes and lung gas volumes compared to animals treated with phospholipids only. This shows that rSP-C33Leu can be purified from bacterial lipids and that rSP-C33Leu surfactant is active against experimental RDS.

Klíčová slova:

Lipids – Phospholipids – Rabbits – Salting out – Surfactants – Tidal volume – Reversed-phase high performance liquid chromatography – Size-exclusion chromatography


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