关键词: 燕窝, 蛋白质, 提取, SDS-聚丙烯酰胺凝胶电泳, 双向电泳

Abstract:

In order to explore the functional components in edible bird’s nest (EBN) and establish a system for the
identification of EBN adulteration and quality classification, two-dimensional electrophoresis (2-DE) was adopted to
separate water-soluble protein from EBN. EBN samples from the Indonesian island of Sumatra were analyzed by 2-DE
and the assay was optimized with respect to extraction solvent, extraction conditions, rehydration buffer composition and
resolving gel concentration. The results showed that 2-DE map of EBN protein with high resolution and good reproducibility
could be obtained under the conditions: 6 h of extraction at 60 ℃ using ultrapure water as extraction solvent, , rehydration
buffer composed of 8 mol/L urea, 4 g/100mL CHAPS, 65 mmol/L DTT, 0.2% Bio-Lyte 4/6 Ampholyte, 0.1% Bio-Lyte 5/8
Ampholyte and 0.001 g/100mL bromophenol blue, and separation gel concentration of 8%. Furthermore, four different EBN
samples were separated successfully by the established 2-DE technology, showing the reliability of the developed method.

Key words: edible-bird&rsquo, s nest；protein；extraction；SDS-polyacrylamide gel electrophoresis (SDS-PAGE)；twodimensional electrophoresis (2-DE)