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Application of Two-Dimensional Electrophoresis Technology in Separation of Water-Soluble Protein from Edible Bird’s Nest

GUO Li-li，WU Ya-jun，LIU Ming-chang，WANG Bin，HAN Jian-xun，GE Yi-qiang，CHEN Ying   

  1. 1. Chinese Academy of Inspection and Quarantine, Beijing 100123, China；
    2. College of Food Science and Nutritional Engineering, China Agricultural University, Beijing 100083, China；
    3. China Rural Technology Development Center, Beijing 100045, China
  • Online:2013-12-25 Published:2014-01-03

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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)