免疫学技术在食品过敏原检测中的应用

食品过敏原可引起机体产生过敏反应,严重危害人类健康及生命安全。目前食品包装上过敏原信息标注规 范尚不完善,因此食品过敏原检测技术对于预防含有过敏原食品进入流通领域,减少过敏事件发生至关重要。本文 综述了免疫学检测技术在食品过敏原检测中应用,并展望了其未来的发展趋势。     

土霉素完全抗原的制备及ELISA检测方法的建立

为制备土霉素(Oxytetracycline,OTC)完全抗原,获得免疫学特性良好的土霉素鼠源多抗血清并建立OTC免疫学检测方法,采用重氮化法在OTC分子上引入羧基,进而利用改进碳二亚胺法将其与载体蛋白偶联制备完全抗原,通过红外扫描、紫外扫描和凝胶电泳鉴定其偶联效果,然后将完全抗原免疫BLAB/c小鼠获得多抗血清(pAb)并对其免疫学特性进行鉴定,通过优化条件建立OTC间接竞争ELISA检测方法。结果表明,偶联效果良好,免疫的4只小鼠多抗血清效价均达到1∶12 800;4只小鼠多抗血清均有抑制效果,其中2号小鼠产生的多抗血清效果最佳,基于该多抗血清建立的OTC间接竞争ELISA检测方法半数抑制浓度(IC_(50))为32.92ng/mL,检测限(LOD)为1.3ng/mL,牛奶样品中添加回收率在84.70%～87.45%,与金霉素、四环素的交叉反应率分别为5.27%,4.10%,与其他竞争物交叉反应率0.4%。该试验成功合成了OTC完全抗原,获得了免疫学特性良好的OTC多抗血清,并建立了OTC免疫学检测方法。     

高灵敏度磁荧光免疫层析试纸模式构建及在呕吐毒素检测中的应用

构建基于磁荧光纳米材料的免疫层析试纸模式,弥补现在免疫层析技术的不足,为更灵敏的免疫学快速检测提供技术支撑。以呕吐毒素（deoxynivalenol,DON）为靶标,采用溶剂热法制备羧基修饰的超顺磁颗粒,碳二亚胺法将磁颗粒、绿色荧光蛋白及DON单克隆抗体进行偶联,一步法制备磁荧光抗体探针,以DON人工抗原（DON-BSA）为检测线建立磁荧光免疫层析试纸。同时用胶体金标记DON单克隆抗体,以DON-BSA为检测线建立胶体金免疫层析试纸;制备的磁荧光抗体探针具有很好的磁性、荧光特性及抗体反应性,基于该探针成功制备了DON磁荧光免疫层析试纸,该试纸回归方程为y=－0.562x＋0.921,R2=0.990,IC50为5.611 ng/mL,检出限为1.089 ng/mL;制备了DON胶体金免疫层析试纸,该试纸裸眼检测灵敏度为500 ng/mL;定量检测回归方程为y=－0.543x＋1.485,R2=0.991,IC50为65.16 ng/mL,检出限为11.94 ng/mL。DON磁荧光免疫层析试纸的灵敏度是胶体金免疫层析试纸的10.96 倍。本实验建立的磁荧光免疫层析试纸模式可以同时实现样品的富集及荧光信号检测,提高检测灵敏度,并成功用于DON的检测,为磁荧光纳米颗粒广泛应用于免疫层析领域提供参考。     

大豆Kunitz胰蛋白酶抑制因子单克隆抗体的制备与鉴定

为制备免疫学特性良好的大豆Kunitz胰蛋白酶抑制因子(Kunitz trypsin inhibitor,KTI)单克隆抗体,以50μg/只的免疫剂量将KTI免疫BALB/c小鼠,利用间接ELISA和间接竞争ELISA鉴定多抗血清效价和敏感性,选择血清效价和敏感性较优的小鼠进行细胞融合,多次筛选得到稳定分泌KTI单克隆抗体(Monoclonal antibody,mAb)的杂交瘤细胞株,采用体内诱生腹水法制备mAb并对其免疫学特性进行鉴定。结果表明,1号小鼠经免疫后效价最高且敏感性最好,半数抑制浓度(IC50)为157.33ng/mL,经细胞融合筛选得到4E6-E9阳性杂交瘤细胞株,所制备的抗体效价达到11 638 400,亚型为IgG1型,亲和常数为1.45×108 L/mol,IC50为28.39ng/mL,且特异性强。说明试验所制备的mAb免疫学特性良好,能够为建立灵敏、特异的KTI免疫学检测方法提供良好的抗体保障。     

化学合成类抗球虫药残留检测方法研究进展

文章综述了化学合成类抗球虫药残留的检测方法,重点介绍了高效液相色谱串联质谱法、酶联免疫吸附测定法、免疫层析试纸法、表面增强拉曼光谱法等的研究进展,总结了检测方法发展现状与趋势,并对其未来发展方向进行了展望.     

大菱鲆疾病早期快速检测方法——胶体金免疫层析试纸的研制与建立

使用成分单一的牛血清白蛋白(BSA)为模拟病原,以胶体金标记兔抗血清(即大菱鲆免疫球蛋白多抗)作为检测示踪物,并分别将BSA和葡萄球菌A蛋白印记到硝酸纤维素膜上制成检测线和对照线,通过一系列工艺创制与组装配套,首次成功制备了一套完整的大菱鲆抗体快速检测试纸。采用大菱鲆抗BSA血清作为阳性样本,以健康大菱鲆血清作为阴性样本,用以检验试纸的性能,并与酶联免疫吸附实验(ELISA)法检测结果相比较。结果表明:本试纸检测抗体的特异性与敏感性均很高,与ELISA方法相当,而且使用方便,不需专业技能和额外的试剂与辅助仪器设备,5 min内即可用裸眼获得观察结果,很适合于基层生产操作及户外调研使用。以该实验为基础建立起来的抗体检测试纸,亦可推广应用于其他病害抗体的检测,可为鱼类疾病早期发生提供简易、快捷和操作性强的诊断方法。     

抗赭曲霉毒素A单克隆抗体的制备及免疫学鉴定

通过碳化二亚胺法(EDC法)制备免疫原OTA-BSA,并免疫小鼠,经杂交瘤技术建立产抗赭曲霉毒素A(OTA)单克隆抗体的杂交瘤细胞株1株。通过体内诱生腹水法制备腹水,纯化后对其亚型、效价、灵敏性、亲和性、特异性等免疫学特性进行鉴定;单抗亚类为IgG1型,间接ELISA效价1:6.4×105,IC50为112.8pg/mL,亲和常数为9.74×1011L/mol,与赭曲霉毒素B交叉反应率为5.6%,与其他常见真菌产物交叉小于0.003%;本实验制备的抗OTA单克隆抗体具有高效、敏感、特异等特点,适合OTA的免疫学快速检测。     

Effect of malt pretreatment on phytic acid and in vitro protein digestibility of sorghum flour

Sorghum seeds of cultivar Wad Ahemed (phytate: 2.7 mg/g, tannin: 0.96% and 2 h pepsin digestion: 18%) were germinated for three days to obtain 1-, 2- and 3- days old sorghum malts. Sorghum malt was added in concentrations of 1, 2.5, 5, 7.5 or 10% to sorghum flour. The mixtures were incubated with shaking for 0, 30, 60, 90 or 120 min. Phytic acid and in vitro protein digestibility were assayed for all treatments. The results revealed that phytate content was significantly reduced. The 10% 3-day-old malt after 120 min incubation, reduced the phytate content by 83%. The in vitro protein digestibility (IVPD) was significantly improved as a result of malt pretreatment. The rate of reduction of phytate content and the rate of increment in IVPD increased with time of incubation, age and concentration of the malt.     

豆芽中生长调节剂类违禁添加物的检测方法研究进展

文章针对近年来频发的"问题豆芽"事件,介绍了豆芽生产过程中禁止添加的6-苄基氨基嘌呤等植物生长调节剂的作用及危害,综述了用于此类生长调节剂检测的高效液相色谱、表面增强拉曼光谱、电化学、免疫分析、生物传感器等方法的应用现状,并对检测方法的发展方向进行展望。     

Synthesis and characterization of a novel fibrous antibacterial fiber with organophorsphor functional groups

A novel antibacterial fiber named ethyltriphenylphosphonium bromide‐polyacrylonitrile fiber (ETPB‐PANF) was synthesized by chemical modification of PANF reacted with ETPB. The PANF was first immersed in NaOH solution to get Na‐PANF with ? COONa groups. Na‐PANF was then reacted with ETPB to get the final fiber. During the process of synthesis, this article investigated on the initial concentration of ETPB, the contact time, the reaction temperature, and the pH of the solution that may have effect on the properties of ETPB‐PANF. ETPB‐PANF was characterized by Fourier transform infrared spectroscopy, thermo gravimetric analyzer, scanning electron microscope, and X‐ray photoelectron spectroscopy, and the releasing amount of ETPB from ETPB‐PANF was examined by inductively coupled plasma atomic emission spectrometry. The antibacterial activity of ETPB‐PANF was examined against Escherichia coli and Staphylococci aureus by improved shake flask method in sterile saline. The results showed that organophorsphor functional groups have been successfully grafted on PANF, and ETPB‐PANF showed good antibacterial abilities for E. coli and S. aureus. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 40935.