Veprek Pavel, Jezek Jan, Velek Jirí, Tallima Hatem, Montash Mona, El Ridi Rashika
Institute of Organic Chemistry and Biochemistry, Academy of Sciences of the Czech Republic, Flemingovo nám. 2, 166 10 Prague 6, Czech Republic.
J Pept Sci. 2004 Jun;10(6):350-62. doi: 10.1002/psc.550.
Four monoepitopic MAPs (MAP A, B, C and E) and one bis-diepitopic MAP B-E derived fromthe primary sequence of Schistosoma mansoni glyceraldehyde 3-phosphate dehydrogenase, previously tested in BALB/c mice, were examined for their immunogenicity and protective capacity in C57BL/6 mice. Despite multimerization into MAPs, MAP Aand MAP C were poorly immunogenic. In contrast toBALB/c mice, MAP E was non-immunogenic in C57BL/6 mice. Peptide B in the form of MAP B orbis-diepitopic MAPB-E elicited immune responses in C57BL/6 mice that were associated with a significant decrease in worm burden. The MAPs were prepared by the stepwise solid-phase peptide synthesis using Boc/Bzl chemistry, successfully purified on the RP-HPLC column and characterized by RP-HPLC, HPCE and MALDI-TOF MS techniques. A general strategy for MAPs purification is discussed here and the purification of MAP Band MAP E is documented in detail.
四种单表位多抗原肽(多抗原肽A、B、C和E)以及一种源自曼氏血吸虫甘油醛-3-磷酸脱氢酶一级序列的双二表位多抗原肽B-E,之前已在BALB/c小鼠中进行过测试,现对其在C57BL/6小鼠中的免疫原性和保护能力进行检测。尽管多抗原肽A和多抗原肽C已多聚化形成多抗原肽,但它们的免疫原性较差。与BALB/c小鼠不同,多抗原肽E在C57BL/6小鼠中无免疫原性。以多抗原肽B或双二表位多抗原肽B-E形式存在的肽B在C57BL/6小鼠中引发了免疫反应,这与虫体负荷的显著降低有关。这些多抗原肽采用Boc/Bzl化学方法通过逐步固相肽合成法制备,在反相高效液相色谱柱上成功纯化,并通过反相高效液相色谱、毛细管电泳和基质辅助激光解吸电离飞行时间质谱技术进行表征。本文讨论了多抗原肽纯化的一般策略,并详细记录了多抗原肽B和多抗原肽E的纯化过程。