| Summary: | 碩士 === 國立中正大學 === 化學工程研究所 === 90 === Abstract
After the human genome has been completely sequenced, the related research of proteomics becomes more important. In this study, the technology of proteomics was used to analyze rabbit serum proteins with hepatic failure and serum proteins from healthy rabbit. The results can be useful to design a bioartificial liver and to help medical doctors for diagnosing the hepatic failure.
In this study, copolymer particles of glycidyl methacrylate (GMA) and styrene were prepared by the method of dispersion copolymerization. The resultant non-porous copolymerized beads were chemically modified to introduce primary amino group, which were ready for the attachment of dye (Cibacron Blue F3GA). The prepared dye-immobilized beads were used to remove albumin from the rabbit serum. After the affinity depletion, we can figure out whether there are other proteins shielded by albumin or not.
In the analysis of two-dimensional gel electrophoresis, we use 17 cm immobilized pH gradient gel (IPG) strips to absorb all proteins in rabbit serum sample. After the electrophoresis procedures of isoelectric focusing and SDS-PAGE, rabbit serum proteins could be well separated according to their pI points and molecular weights.
After the silver stain, the two-dimensional gel was scanned by using an image scanner and protein spots were analyzed by the PDquest software. Comparing serum proteins from diseased rabbit with those from healthy one, some marker proteins that changed with hepatic failure were asserted. The marker spots in the two-dimensional get were cut, digested with trypsin, and analyzed by MALDI-TOF mass spectrometry.
Matching their peptide mass fingerprints with database identified four marker proteins. They are Serum amyloid A-3 protein precursor (SAA3), Serum amyloid A-2 protein precursor (SAA2), Hypothetical protein and Putative serum amyloid A protein. SAA3 and SAA2 are members of the SAA family of acute-phase or inflammatory response proteins. Results obtained in this work show that the expression level of these proteins decreased, or even disappeared, during the process of hepatic failure.
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