Abstract
The surfactant proteins (SPs), SP-B and SP-C, are important components of pulmonary surfactant involved in the reduction of alveolar surface tension. Quantification of SP-B and SP-C in surfactant drugs is informative for their quality control and the evaluation of their biological activity. Western blot analysis enabled the quantification of SP-B, but not SP-C, in surfactant drugs. Here, we report a new procedure involving chemical treatments and LC-MS to analyze SP-C peptides. The procedure enabled qualitative analysis of SP-C from different species with discrimination of the palmitoylation status and the artificial modifications that occur during handling and/or storage. In addition, the method can be used to estimate the total amount of SP-C in pulmonary surfactant drugs. The strategy described here might serve as a prototype to establish analytical methods for peptides that are extremely hydrophobic and behave like lipids. The new method provides an easy measurement of SP-C from various biological samples, which will help the characterization of various experimental animal models and the quality control of surfactant drugs, as well as diagnostics of human samples.
Highlights
The surfactant proteins (SPs), SP-B and SP-C, are important components of pulmonary surfactant involved in the reduction of alveolar surface tension
SP-B and SP-C are hydrophobic peptides that act in concert with the major pulmonary surfactant phospholipid, dipalmitoylphosphatidylcholine (DPPC), and contribute to the surface tension-lowering property
The initial purpose of our study was the quantification of SP-B and SP-C in a surfactant drug, Surfacten
Summary
The surfactant proteins (SPs), SP-B and SP-C, are important components of pulmonary surfactant involved in the reduction of alveolar surface tension. Quantification of SP-B and SP-C in surfactant drugs is informative for their quality control and the evaluation of their biological activity. Western blot analysis enabled the quantification of SP-B, but not SP-C, in surfactant drugs. The new method provides an easy measurement of SP-C from various biological samples, which will help the characterization of various experimental animal models and the quality control of surfactant drugs, as well as diagnostics of human samples.—Harayama, T., H. Pulmonary surfactant is a surface-active material secreted in the alveolar space. It reduces alveolar surface tension upon compression and prevents alveolar collapse at end-expiration. SP-B and SP-C are hydrophobic peptides that act in concert with the major pulmonary surfactant phospholipid, dipalmitoylphosphatidylcholine (DPPC), and contribute to the surface tension-lowering property.
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