酶联免疫吸附分析法检测花生过敏原的研究

本文利用自己研制的花生过敏原免疫新西兰大耳兔,获得效价为200 000的抗花生过敏原特异性抗体,建立了花生过敏原蛋白的间接竞争酶联免疫(ELISA)检测方法.结果表明:花生抗原在0.01～100 ng/mL范围内具有较好的线性关系,其竞争标准曲线为v=-0.1212x+0.9285(r=0.9819),IC50为34....     

非标记型河虾免疫传感器的制备及应用

四乙氧基硅在HCl催化下水解形成硅溶胶。将硅溶胶与河虾抗体混合均匀后,涂于玻碳电极表面制备得非标记型河虾抗体免疫传感器。采用Fe(CN)63-/4-的磷酸盐缓冲溶液(pH 6.5)为测试底液,研究此传感器在免疫反应中的循环伏安和交流阻抗特性。结果显示免疫反应后传感器的循环伏安图上未出现新的氧化-还原峰,表明该免疫反应属于非氧化还原过程。采用交流阻抗监测电极表面,发现传感器阻抗值随虾过敏原浓度的增加而增大。对交流阻抗图进行分析,结果表明电极表面的免疫反应是一个受电子转移控制的过程。当虾过敏原浓度在0~10 ng/mL之间,电极表面电子转移阻抗的增加值与虾过敏原浓度呈线性关系,检出限为0.1 ng/mL(S/N=3)。该方法已成功应用于河虾制品中过敏原的测定。     

胶体金免疫层析方法快速检测腹泻性贝毒软海绵酸的初步研究

建立了赤潮毒素腹泻性贝毒软海绵酸的快速胶体金免疫层析检测方法。通过细胞融合,制备抗软海绵酸单克隆抗体,胶体金标记抗体,建立快速检测软海绵酸的免疫层析试纸条方法。检出限500 ng/mL(50 ng/条),探讨了影响测试方法的因素和提高灵敏度的可能手段。     

氟苯尼考胶体金免疫层析定量检测方法的建立

建立了定量检测氟苯尼考的胶体金免疫层析方法.对胶体金标记抗体时溶液pH和抗体浓度、金标抗体用量、检测线上抗原浓度以及检测时间进行了优化.采用胶体金试纸条读取仪测定试纸条检测线和质控线的信号强度,以标准品的浓度为横坐标,阳性样本和阴性样本的检测线/质控线的信号比值(Bx/B0)为纵坐标建立标准曲线.结果表明,胶体金免疫层析试纸定量检测氟苯尼考的线性范围为0.1~1.5 ng/mL,检出限为0.08 ng/mL,检测时间为15 min.本方法具有简便、快速和可定量等特点,适于大批量样品的现场筛查.     

胶体金免疫层析法快速检测水产品中河豚毒素的研究

该文将特异性识别河豚毒素的单克隆抗体加以胶体金标记用作示踪物,建立了豚毒素的胶体金免疫层析技术快速检测方法。优化了胶体金体系的pH值、抗原抗体浓度、离子浓度、表面活性剂种类以及样品前处理方法。结果显示：在最优条件下,建立的胶体金免疫层析法对河豚毒素的定量检出限为0.5 ng/mL,线性范围为0.8～10.6 ng/mL,定性检出限（裸眼判别）为12.0 ng/mL。河豚、织纹螺等样品的加标回收率为70.5%～110%,相对标准偏差为3.7%～7.1%,检测结果与LC-MS/MS法一致。所建立的胶体金免疫层析技术在现场快速检测方面具有良好的可行性和实用性,可用于大量样品的快速筛查和河豚毒素中毒后的快速诊断。     

Preparation and Characterization of Olaquindox Polyclonal Antibody

本研究先将喹乙醇琥珀酸单酯化,使其转变为带有羧基的衍生物。合成产物经重结晶纯化,得率为52.47%。合成产物进行结构鉴定,合成了具有喹乙醇分子结构特征的喹乙醇半抗原。采用活化酯法将半抗原与牛血清白蛋白（BSA）进行偶联,采用混合酸酐法将半抗原与卵清蛋白（OVA）进行偶联,分别制备免疫原和包被抗原。紫外扫描与红外图谱分析结果表明,半抗原与载体蛋白偶联成功,与BSA、OVA的结合比分别为3.8:1和5.0:1。以OLA-BSA作为免疫原免疫4只新西兰大白兔,获得了较高效价的抗血清,以OLA-OVA作为包被抗原,间接ELISA法测定各兔的效价,分别为1:6400;1:1600;1:12800;1:6400。间接ELISA法喹乙醇的抑制中浓度IC50为743.3 ng/mL,最低检测限IC20为5.71 ng/mL。从电泳图谱分析可以看出,经纯化得到了纯度较高的喹乙醇多克隆抗体,为喹乙醇免疫分析方法的进一步研制和开发奠定基础。     

基于酶催化沉积质量放大的压电免疫传感器的研究

提出了一种酶催化沉积质量放大的高灵敏压电免疫传感器. 采用1,6-二巯基己烷在石英晶振上自组装一单分子层, 再通过另一端巯基连接胶体金, 利用胶体金的高比表面积和强吸附作用力增加抗IgG抗体的固定量, 同时借助胶体金优良的生物亲和性保持抗IgG抗体的活性. 在H2O2存在下, 通过标记在抗人IgG抗体上的HRP酶催化底物DAB(3,3′-联苯二胺), 反应中生成的不溶性产物沉积到石英晶振的Au电极表面, 达到质量放大的目的. 结果表明, 检测人IgG在16 ng/mL-100 μg/mL范围内有很好的线性关系, 检测下限为10 ng/mL, 在用于实际试样的回收率测定中, 结果良好.     

微孔板蛋白芯片可视化检测方法的研究

将微孔板生物芯片制备技术与银增强显色方法相结合,建立了一种高通量蛋白免疫分析新方法。研究采用夹心法反应原理,将人IgG捕获抗体以微阵列形式点样于96孔板底部制备成蛋白微阵列,并依次与人IgG、生物素标记二抗及链亲和素胶体金反应,最后用银增强法显色。采用平板式扫描仪对微孔板显色结果进行快速扫描成像,图像采用luxscan 3.0软件处理。结果表明:微孔板蛋白芯片的最低检测量可达0.6 ng/mL,线性范围10 ng/mL~100μg/mL,相关系数为0.98,重复性较好(CV10%)。     

莱克多巴胺免疫亲和柱的制备与应用研究

用多元酸酐与混合酸酐相结合的方法合成了莱克多巴胺(Rac)抗原,免疫动物获得特异性抗体,并以蛋白A柱纯化得到IgG抗体。琼脂糖凝胶(Sepharose 4B)经溴化氰(CNBr)活化后与IgG抗体偶联,制备莱克多巴胺免疫吸附剂。据此建立了尿液中莱克多巴胺的免疫亲和柱净化/液相色谱-荧光法(HPLC-FL)测定的分析方法。免疫制备特异抗体50%抑制浓度(IC50)为5μg/L。Sepharose 4B经CNBr活化后与2 mg抗体的偶联率达87.4%。1 mL吸附剂的柱容量为67.57 ng。尿液中莱克多巴胺的回收率为76%~90%。     

粮食中百草枯残留的金标免疫层析检测方法研究

采用柠檬酸三钠还原法制备胶体金,研究胶体金与抗体蛋白的作用过程,确定稳定标记1.0 mL胶体金需8.10μg抗体蛋白,标记体系的最佳pH值为8.5。以金标抗体为分析探针,PQ-h-OVA为竞争抗原,羊抗兔IgG为控制抗体,构建直接竞争胶体金标免疫层析检测体系(GICA)。确定膜上包被抗原的质量浓度为0.50 g/L,点样量为1.0μL/条;金标点样量:5.0μL/条;羊抗兔二抗的最佳包被质量浓度为0.11 g/L,点样量为1.0μL/条。金标试纸条的目测检出限为10μg/L,检测时间约5 min,交叉反应率小于0.10%。方法的重复性和稳定性较好,该试纸条在室温下至少可保存5个月。     

Furyl- and thienyl-silatranes and germatranes

We review our research on the synthesis and study of the physical and biological properties of furyl- and thienylgermatranes and -silatranes.Translated from Khimiya Geterotsiklicheskikh Soedinenii, No. 6, pp. 725–732, June, 1992.     

Review and patents and literature

The use of the insect cell/baculovirus expression system for producing recombinant proteins of bacterial, plant, insect, and mammalian origin has become widespread. The popularity of this eukaryotic expression system is due to many factors, including (1) potentially high protein expression levels, (2) ease and speed of genetic engineering, (3) ability to accommodate large DNA inserts, (4) protein processing similar to higher eukaryotic cells (e.g., mammalian cells), and (5) ease of insect cell growth (e.g., suspension growth). The following review of the literature discusses two engineering aspects of recombinant protein synthesis by insect cell cultures: bioreactor scale-up and insect cell line selection. Following this review patent abstracts and additional literature pertaining to expression of recombinant proteins in insect cell culture are listed.     

多环芳二酐型聚酯亚胺膜的透气性能

多环芳二酐型聚酯亚胺膜的透气性能李悦生，丁孟贤，徐纪平（浙江大学高分子科学与工程研究所，杭州，３１００２７）（中国科学院长春应用化学研究所）关键词聚醚酰亚胺，聚酯酰亚胺，膜，透气性通常的聚酰亚胺加工性能较差，在芳环二酐的苯环间引入醚键等柔性基团后，其...     

微量元素与疾病诊断治疗的研究现状及展望

对国内外微量元素与疾病的关系及其诊断治疗的研究现状及研究动向进行了综述，包括：微量元素与地方病、癌肿、心血管病、糖尿病、眼病、神经系统病等。参考文献72篇。     

Preparation and photolysis of poly-p-ethylacrylophenone and poly-p-chloroacrylophenone and their copolymers with styrene and methyl methacrylate

Poly-p-ethylacrylophenone (PPEtAP) and poly-p-chloroacrylophenone (PPClAP) and their copolymers with styrene (PEtAP/S) and (PClAP/S) and with methyl methacrylate (PEtAP/MMA) and (PClAP/MMA) were prepared. Quantum yields of main-chain scissions at 366 nm at room temperature in benzene solution are of the same order as those of the unsubstituted polyacrylophenone and its copolymers. Substitution with chlorine and ethyl group in the para position compared to the unsubstituted polyacrylophenone and its copolymers leads to an increase of the lifetime for the n–π^* excited triplet state, as is evident from the measured quenching constants of photolysis. The low-temperature emission spectra of the copolymers are similar to the spectra of the model compounds in a polymer film. For the homopolymer, however, the character of the emission spectrum changes considerably.     

Concentrations and distributions of trace and minor elements in Chinese and Canadian coals and ashes

A total of 35 trace and minor elements including some of environmental significance were determined in each of a selection of 15 Chinese and 6 Canadian thermal coals and their ashes by using the SLOWPOKE-2 nuclear reactor facility of the University of Toronto. The concentrations and distributions of these constituents among the coals and their combustion products (viz. ash and volatile matter) are presented together with an interpretation of their significance in relation to the large scale combustion of these coals as thermal fuels in industrialized countries such as China and Canada. The detailed results showed wide variations in trace impurity concentrations (up to a factor of 100 and more) among the coals studied with few large differences between those of Chinese and Canadian origin except that the rare earths, Sc, Th, U, I, and Se were much higher in the former, other halogens, As and Na were lower. Values for elemental enrichment factors (EF) relative to normal crustal abundances indicated that only As(EF=13), Br(5.7), I(16), S(230), Sb(11) and Se(320) were appreciably enriched in coal. During static ashing at 750°C most of the halogens, S and Se were volatilized whereas most other inorganic constituents were highly retained and concentrated in the ash by factors of 6 to 11.     

Ground- and excited-state aromaticity and antiaromaticity in benzene and cyclobutadiene

The aromaticity and antiaromaticity of the ground state (S 0), lowest triplet state (T 1), and first singlet excited state (S 1) of benzene, and the ground states (S 0), lowest triplet states (T 1), and the first and second singlet excited states (S 1 and S 2) of square and rectangular cyclobutadiene are assessed using various magnetic criteria including nucleus-independent chemical shifts (NICS), proton shieldings, and magnetic susceptibilities calculated using complete-active-space self-consistent field (CASSCF) wave functions constructed from gauge-including atomic orbitals (GIAOs). These magnetic criteria strongly suggest that, in contrast to the well-known aromaticity of the S 0 state of benzene, the T 1 and S 1 states of this molecule are antiaromatic. In square cyclobutadiene, which is shown to be considerably more antiaromatic than rectangular cyclobutadiene, the magnetic properties of the T 1 and S 1 states allow these to be classified as aromatic. According to the computed magnetic criteria, the T 1 state of rectangular cyclobutadiene is still aromatic, but the S 1 state is antiaromatic, just as the S 2 state of square cyclobutadiene; the S 2 state of rectangular cyclobutadiene is nonaromatic. The results demonstrate that the well-known "triplet aromaticity" of cyclic conjugated hydrocarbons represents a particular case of a broader concept of excited-state aromaticity and antiaromaticity. It is shown that while electronic excitation may lead to increased nuclear shieldings in certain low-lying electronic states, in general its main effect can be expected to be nuclear deshielding, which can be substantial for heavier nuclei.