中文摘要：

目的人工合成N-酰基高丝氨酸内酯（N-acylated homoserine lactones,N-aHSLs）完全抗原并制备相应单抗,建立N-aHSLs免疫检测的新方法及海洋弧菌N-aHSLs动态监测。方法以月桂酸和高丝氨酸内酯为主要原料,通过多步系列化学反应人工合成N-十二酰高丝氨酸内酯（N-lauroyl-homoserine lactone,N-C（12）-HSL）及含羧基活性交联基团的半抗原衍生物12-羧基-N-十二酰高丝氨酸内酯（12-carboxyl-N-decanoyl-L-homoserine lactone,12-CD-LHL）,用1H-磁共振、高压/效液相色谱仪和液相色谱-质谱联用方法确证合成产物的结构,并用生物感应法检测合成N-aHSLs的天然生物活性;用N-羟基琥珀酰亚胺活泼酯法将12-CD-LHL半抗原衍生物连接到载体蛋白;将N-C（12）-HSL人工完全抗原通过传统单抗制备技术,获得抗N-aHSLs杂交瘤细胞株,用酶联免疫吸附测定法鉴定其特异性和效价。结果质谱、磁共振氢谱分析结果与理论预测相同,成功合成目标产物N-C（12）-HSL和12-CD-LHL半抗原衍生物;所合成的N-aHSLs能够分别激活生物感应菌株大肠杆菌MG4和紫色色杆菌026并使之显色;紫外吸收光谱扫描结果显示,半抗原已分别与牛血清白蛋白和卵清蛋白发生偶联,成功制备完全抗原;共获得3株抗N-C（12）-HSL单抗,均具有良好的免疫反应性。结论 N-aHSLs完全抗原和相应的单抗制备为N-aHSLs动态监测和后续研究奠定了基础。

英文摘要：

Objective To synthesize the artificial holoantigen and corresponding monoclonal antibodies of N-acylated homoserine lactones（N-aHSLs）,and to provide basis for establishment of a new immunodetection and monitoring method of N-aHSLs.Methods Lauric acid and homoserine lactone were used as the starting materials to synthesize N-lauroyl-homoserine lactone（N-C（12）-HSL）and the hapten derivant of 12-carboxyl-N-decanoyl-L-homoserine lactone（12-CD-LHL） with chemical crosslinking active group by serial reactions.The target products were characterized by1 H nuclear magnetic resonance（1H-NMR）,high pressure/performance liquid chromatography（HPLC）and liquid chromatograph-mass spectrometer（LC-MS）.The natural bioactivity of N-aHSLs was detected by biological response method.Holoantigen of N-C（12）-HSL was synthesized by the method of active ester.BALB/c mice was immunized with N-C（12）-HSL holoantigen,and hybridoma cells against N-C（12）-HSL were produced by cell-fusion technique.The specificity and titers of monoclonal antibody against N-C（12）-HSL were screened by enzyme-linked immunosorbent assay.Results The1H-NMR,HPLC and LC-MS results revealed that the obtained compound were the target product of N-C（12）-HSL and 12-CD-LHL,just as the prediction.The synthetical N-aHSLs could activate the biological response strains of escherichia coli and chromobacterium violaceum 026 seperately and color the strains.The ultraviolet absorption spectrum results revealed that hapten N-C（12）-HSL had crosslinked with bovine serum albumin and ovalbumin,N-C（12）-HSL holoantigen had been prepared well.Three strains of hybridoma against N-C（12）-HSL were obtained,and the identification results indicated which all have favourable immunoreactivity.Conclusion The preparation of N-aHSLs whole antigen and corresponding monoclonal antibody laid the foundation for N-aHSLs dynamic monitoring and subsequent research.