Comparison of antigenic proteins between pathogenic E. Coli by employing immunoproteomics
Justiniane, Ferliza E
전기영동 병원성대장균 proteom;
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Escherichia coli is a gram-negative bacterium that causes serious diseases, particularly in digestive and urinary tract of humans and animals. In order to determine and compare antigenic proteins between 8 strains of pathogenic E. coli, proteomics technology with the combination of Matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF MS) and immunoblot analysis were utilized. Proteins were separated in the first dimension by Isoelectric focusing (IEF) using 13 cm immobilized pH gradient (IPG) strips with pH 4-7. This was followed by protein loading at 12.5% gradient Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Results revealed that a total of 69 antigenic spots out of 182 and corresponds to 42 proteins were determined in strain ACH5 using E. coli O157:H7 antiserum, 87 of 182 spots corresponding to 43 proteins in K88, 86 of 154 spots that corresponds to 30 proteins in K99, 91 of 210 spots in O148 which corresponds to 14 proteins, 50 of 222 spots that corresponds to 10 proteins in O59, 38 proteins corresponded to 75 of 247 spots in O157:H7, 83 of 286 spots which corresponds to 22 proteins in C2, and 67 antigenic spots out of 156 that corresponds to 42 proteins were determined in O26. On the other hand, strain specific antisera detected a total of 46 out of 102 spots in K88 strain, 10 of 31 spots in K99, 18 of 35 spots in O148, 75 of 247 spots in O157:H7 and 29 of 41 spots in O26. Common as well as specific antigenic proteins were being noted and differentiated between respective strains. Comparison conducted reveals a total of 15 common antigenic spots present in each strain as recognized by E. coli O157:H7 antiserum. In contrast, specific antigenic proteins identified differ in total numbers between strains. E. coli K88 and O148 were identified having 17 specific antigenic spots, a total of 10 in strains K99 and O59, 7 in E. coli C2, O1578:H7 and O26 strains, whereas 4 were present in ACH5. Data gathered indicate that greater similarities observed in 2-DE profiles suggests the use of broad spectrum antibodies that effectively recognize antigenic proteins of pathogenic E. coli. Furthermore, protein differences noted might be due to variations between host and their degree of pathogenicity. Antigenic proteins identified would be useful in employing either vaccine development or diagnostic marker, or both and would be applicable for epidemiological studies.