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诺氟沙星与牛血清白蛋白及卵清蛋白结合物的合成 总被引:7,自引:0,他引:7
采用碳二亚胺法制备诺氟沙星与牛血清白蛋白(BSA)的结合物,作为诺氟沙星酶联免疫用的免疫原。经紫外光谱法测定,每分子牛血清白蛋白连接的诺氟沙星分子数为16.1个。分别用碳二亚胺法和混合酸酐法制备诺氟沙星与卵清蛋白(OA)的结合物,作为诺氟沙星酶联免疫测定用的包被抗原。经紫外光谱法测定,每分子卵清蛋白连接的诺氟沙星分子数分别为5.8和1.9个。 相似文献
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采用碳二亚胺法制备诺氟沙星与牛血清白蛋白(BSA)的结合物,作为诺氟沙星酶联免疫用的免疫原。经紫外光谱法测定,每分子牛血清白蛋白连接的诺氟沙星分子数为16.1个。分别用碳二亚胺法和混合酸酐法制备诺氟沙星与卵清蛋白(OA)的结合物,作为诺氟沙星酶联免疫测定用的包被抗原。经紫外光谱法测定,每分子卵清蛋白连接的诺氟沙星分子数分别为5.8和1.9个。 相似文献
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采用N-羟基琥珀酰亚胺活性酯法,将恩诺沙星(ENR)分别与牛血清白蛋白(BSA)和卵清蛋白(OVA)连接,制备人工免疫原ENR-BSA和包被原ENR-OVA,经紫外扫描分析和动物免疫试验证实人工抗原合成成功,研究结果对抗恩诺沙星单克隆抗体的制备具有重要意义. 相似文献
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为了制备诺氟沙星(NFLX)抗体,将修饰成功的NFLX-NH2分别与牛血清白蛋白(BSA)和卵清蛋白(OVA)偶联,制备完全抗原,用免疫抗原免疫小鼠,获得了高效价的诺氟沙星多克隆抗体. 相似文献
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【目的】制备培氟沙星(Pefloxacin,PEF)完全抗原,并对其免疫学特性进行鉴定。【方法】以培氟沙星与牛血清白蛋白(BSA)为材料,用碳二亚胺(EDC)法制备培氟沙星-牛血清白蛋白完全抗原(PEF-BSA),变性聚丙烯酰胺凝胶电泳(SDS-PAGE)及紫外扫描(UV)检测其免疫学特性;利用PEF-BSA免疫BALB/c小鼠,无菌条件下收集血清,用间接ELISA测定多抗血清(pAb)效价。并用同样的方法制备和鉴定培氟沙星-鸡卵清白蛋白完全抗原(PEF-OVA)。【结果】培氟沙星与牛血清白蛋白及鸡卵清白蛋白偶联成功,PEF-BSA偶联比为8∶1,PEF-OVA偶联比为6∶1。间接ELISA测得多抗血清效价均达到4 000以上,得到了较好的免疫效果。【结论】成功得到了培氟沙星完全抗原,该抗原具有较强的特异性及良好的敏感性。 相似文献
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为了制备环丙沙星(CPLX)多克隆抗体,将修饰成功的CPLX-NH2分别与牛血清白蛋白(BSA)和卵清蛋白(OVA)偶联,制备完全抗原—CPLX-NH2-BSA和CPLX-NH2-OVA,用CPLX-NH2-BSA免疫小鼠,获得了高效价的环丙沙星多克隆抗体. 相似文献
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采用N-羟基琥珀酰亚胺活性酯法,将恩诺沙星(ENR)分别与牛血清白蛋白(BSA)和卵清蛋白(OVA)连接,制备人工免疫原ENR-BSA和包被原ENR-OVA,经紫外扫描分析和动物免疫试验证实人工抗原合成成功,研究结果对抗恩诺沙星单克隆抗体的制备具有重要意义. 相似文献
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An ester activation method was employed to couple enrofloxacin(ENFX) to the carrier proteins BSA and OVA. The conjugates ENFX-BSA and ENFX-OVA were identified with an UV spectrophotometer and amino acid automation analysis instrument, and resulted in conjugates with 48 ENFX molecules per carrier molecule(BSA). Splenocytes from mice immunized with ENFX-BSA were fused with SP2/0 myeloma cells and hybridomas secreting antibodies against enrofloxacin were selected and cloned. Two stable monoclonal antibodies, 2C5, 5D5 of the subclass IgG2a, were isolated. Using antibody 5D5, an indirect competitive inhibition enzyme-linked immunosorbent assay (Ci-ELISA) was developed for the quantitative detection of enrofloxacin and its metabolites. The IC50 of the standard curve was 21.67 ng mL-1 and the limit of detection for enrofloxacin was 0.13 ng mL-1. This method was sensitive and had a linear range from 0.13 to 10 000 ngmL-1 (r=-0.9782). Monoclonal antibody 5D5 exhibited high relative affinity to enrofloxacin, and the cross-reactivities with ciprofloxacin,marbofloxacin, sarafloxacin and danorfloxacin were 110.8, 27.40, 71.05 and 37.41%,respectively. Three non-fluoroquinolones of cefadroxil, chloramphenicol, sulfadimethoxine were tested and there was no cross-reaction between them. 相似文献
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采用碳二亚胺法将金霉素(chlortetracyline,CTC)偶联到载体蛋白BSA(牛血清白蛋白)和OVA(鸡卵清蛋白)上,形成免疫原BSA-CTC与包被原OVA-CTC。通过紫外分光光度法和聚丙烯酰胺凝胶电泳法进行抗原鉴定,用间接ELISA测定多抗血清(pAb)效价,阻断ELISA鉴定其敏感性。结果表明,在UV下出现了明显特征峰,BSA-CTC的峰与BSA、CTC相比均发生改变,表明半抗原与载体成功偶联。共免疫了5只小鼠,pAb效价均达到1×10-3,其中3号小鼠多抗血清抑制效价最低,IC50(半数抑制质量浓度)为71.47ng/mL。由此得出,获得了高效价、强特异的盐酸金霉素多抗血清,为盐酸金霉素单抗的制备奠定了基础。 相似文献
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吡虫啉人工抗原的合成与鉴定 总被引:8,自引:2,他引:8
以吡虫啉原药与β-巯基丙酸为起始原料,在碱性条件下反应,合成了半抗原{1-[6-(2-羧基乙硫基)-3-吡啶基甲基]-N-硝基亚咪唑烷-2-叉胺}。利用该半抗原与牛血清蛋白(BSA)和卵清白蛋白(OVA)偶联得到了人工抗原,经免疫动物后,获得了高效价的特异性多克隆抗体,抗血清效价为2.56×104。经酶联免疫反应(ELISA)测定,吡虫啉对抗体的抑制中浓度(IC50)为 20.7 ?g·L-1,最低检测限(IC20 )为1.1 ?g·L-1,检测范围在1~1 000 ?g·L-1内线性关系较好,吡虫啉结构类似物交叉反应率均小于1.4%。 相似文献
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Hapten sulfaguanidine (SG) was coupled with carrier protein bovine serum albumin (BSA) to form a full antigen SG- BSA by diazotization and glutaraldehyde methods. Ovalbumin (OVA) was used as protein carrier to couple with Hapten SG by glutaraldehyde method. Then, the immunogen and the coating antigen were purified by dialysis and gel exclusion chromatography. The conjugated ratio of SG to BSA in artificial antigen was 5.3 (using diazotization method) and 6.5 (glutaraldehyde method), and the conjugated ratio of SG to OVA in coating antigen was 2.3 (glutaraldehyde method) by UV-visible spectrophotometer. The coupling was successful according to the analysis of sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). BALB/c mice were immunized with the antigen (SG-BSA), and the titers of antiserum were tested to be 1 : 6 400 and 1 : 400 after three periods of immunities by indirect ELISA, which further identified the success of the synthesis of both immunogen SG-BSA and coating antigen SG-OVA. 相似文献