抗黄曲霉毒素B1重链IgG2b亚型单克隆抗体的制备及应用

目的制备抗黄曲霉毒素B1(AFB1)的重链IgG2b亚型单克隆抗体,并建立黄曲霉毒素B1的间接竞争ELISA检测方法。方法采用AFB1-BSA偶联物为免疫抗原,AFB1-STI偶联物为检测抗原,利用间接竞争ELISA筛选分泌抗AFB1单抗的细胞株,并对抗体进行鉴定。结果筛选到1株能分泌特异性抗AFB1单克隆抗体的杂交瘤细胞株,其染色体数目为(90±10)条;所分泌的单抗亚类重链为IgG2b,轻链为λ;间接ELISA检测该株杂交瘤细胞分泌上清效价在1∶12800～1∶25600,纯化腹水效价为1∶105～1∶106。传30代及液氮中保存6个月,抗体效价稳定;建立的间接竞争ELISA检测方法的最低检出限为0.001ng/ml,校正曲线的线性范围为0.005～5ng/ml,IC50为0.01ng/ml;与AFB1的结构类似物AFM1和化学结构有差异的脱氧雪腐镰刀菌烯醇的交叉反应率分别为1.4%和<1%。结论所制备的抗黄曲霉毒素B1单克隆抗体具有高度特异性和稳定性,可应用于间接竞争ELISA检测方法。     

玉米赤霉烯酮单克隆抗体的制备及鉴定

采用玉米赤霉烯酮(ZEN)与羧甲基羟胺半盐酸盐在吡啶中反应,生成玉米赤霉烯酮肟(ZENO),用活性酯法将ZENO与牛血清白蛋白(BSA)的氨基相连合成ZEN人工抗原;并进行了紫外吸收鉴定;以合成的BSA偶联物(ZEN-BSA)为免疫抗原,免疫Balb/c小鼠,经细胞融合,采用非竞争/竞争ELISA两步筛选法获得能够稳定分泌抗体的杂交瘤细胞株1C7.抗体1C7灵敏度(IC50)达0.61μg/L,与T-2毒素、呕吐毒素(DON)、黄曲霉毒素(AFT)的交叉反应率均＜0.1％.本研究为研发粮油产品中玉米赤霉烯酮特异性免疫分析技术及产品奠定了重要基础.     

溴氰菊酯人工抗原及多克隆抗体制备

以二溴菊酸和4-[(4-硝基苯)氧基[苯甲醛为原料经4步反应合成了溴氰菊酯半抗原1-氰基[4-(4-氨基苯)苯基]甲基-2,2-二甲基3-(2',2'-二溴乙烯基)环丙烷羧酸酯;用重氮化法将溴氰菊酯半抗原分别与牛血清白蛋白(BSA)和卵清白蛋白(OVA)相偶联制备免疫抗原和包被抗原,紫外吸收法估算偶联比分别为11:l和8:1;以BSA偶联物作免疫抗原免疫日本大耳白兔制备溴氰菊酯多克隆抗体,间接非竞争ELISA法测定两个抗血清效价分别为100000和80000,间接竞争ELISA法测定抗体特异性结果表明所制备的抗体测定溴氰菊酯的IC50值为0.16 mg/L,与其他供试农药的交叉反应率均≤1%.制备的溴氰菊酯人工抗原和多克隆抗体为研究建立溴氰菊酯农残免疫快速测定方法奠定了基础.     

三聚氰胺完全抗原的制备与表征

采用戊二醛法将三聚氰胺(Melamine,MEL)偶联到牛血清白蛋白(BSA)上制备完全抗原MEL-BSA,并用紫外扫描、红外扫描及SDS-PAGE电泳试验进行了鉴定。结果表明,半抗原MEL和BSA偶联成功,紫外光谱分析法计算得其偶联比为7.9∶1。通过免疫动物获得效价为1∶25 600的抗体,间接竞争ELISA测定抗体IC50值为39 ng/mL,与三聚氰酸及其他几种常用抗菌素的交叉反应率均0.2%,为下一步研制三聚氰氨单克隆抗体,建立三聚氰氨免疫速测技术奠定了基础。     

氰戊菊酯免疫分析关键试剂人工抗原的合成与应用效果

以氰戊菊酸、间苯氧基苯甲醛为原料,利用酰氯与伯胺的活泼反应,合成了一种含有3个碳原子长度连接臂的氰戊菊酯半抗原,并通过核磁共振鉴定;用活泼酯法将合成的氰戊菊酯半抗原分别与牛血清白蛋白(BSA)和卵清白蛋白(OVA)偶联制备免疫抗原和检测抗原,紫外-可见连续光谱扫描结果显示,所制备的人工抗原的图谱具有载体蛋白和半抗原的特征峰叠加现象,表明偶联成功;人工抗原的免疫结果显示,以BSA偶联物免疫小鼠后获得抗血清效价分别为160 000、140 000和110 000,间接竞争ELISA(酶联免疫吸附分析)测定抗体IC50(半数抑制浓度)值为35.1 μg/L,与其他供试农药的交叉反应率均＜1%,为氰戊菊酯抗体研制及免疫分析检测技术研究提供了关键试剂.     

呋喃妥因代谢物AHD单克隆抗体的制备及鉴定

目的制备呋喃妥因代谢物AHD单克隆抗体,并进行鉴定。方法利用对羧基苯甲醛合成1-氨基乙内酰胺脲(AHD)的衍生物1-氨基乙内酰脲-4-羧苯基肟(4-CPAHD),通过活性脂法将4-CPAHD与牛血清白蛋白(BSA)和卵清蛋白(OVA)偶联,获得免疫抗原4-CPAHD-BSA和包被抗原4-CPAHD-OVA,采用紫外分光光度法检测偶联是否成功。将人工合成抗原4-CPAHD-BSA免疫BALB/c小鼠,通过杂交瘤技术将免疫小鼠脾脏细胞与Sp2/0骨髓瘤细胞融合,筛选能稳定产生抗体的细胞株;通过间接ELISA法检测该细胞株分泌的单克隆抗体的效价、邻硝基-PAHD(2-NPAHD)对单克隆抗体的半数抑制浓度(IC50),并分析抗体的特异性。结果偶联后4-CPAHD-BSA的最大吸收峰有较明显的偏移,表明4-CPAHD与BSA偶联成功,平均偶联比为17.4∶1。制备的单克隆抗体的效价为1∶64 000,2-NPAHD对单克隆抗体的IC50为1.45μg/L,单克隆抗体与同类抗生素及其代谢物无交叉反应。结论制备的AHD单克隆抗体各项指标均较好,为呋喃妥因代谢物免疫检测试剂的研发奠定了基础。     

玉米赤霉烯酮抗独特型单抗的制备及特异性分析

目的制备并鉴定玉米赤霉烯酮(Zearalenone,ZEN)抗独特型单抗Ab2β-1D5。方法将ZEN单抗1G4与载体蛋白KLH偶联作为免疫原,免疫BALB/c小鼠,取免疫小鼠脾细胞,与小鼠骨髓瘤细胞SP2/0融合后,以ZEN单抗Fab片段作为包被抗原,间接ELISA法筛选阳性杂交瘤细胞。经小鼠腹腔注射杂交瘤细胞,制备腹水型单抗,并经Protein G亲和层析柱进行纯化。间接ELISA法检测ZEN抗独特型单抗的抗体效价及特异性;间接竞争ELISA法检测ZEN抗独特型单抗类型、灵敏度及其与ZEN毒素间的相关性。结果共获得6株稳定分泌ZEN抗独特型单抗的杂交瘤细胞株,腹水型抗体效价为1∶1.2×105～1∶2.0×105;6株单抗均为β型抗独特型抗体(Ab2β),其中Ab2β-1D5抗体灵敏度最高,对ZEN的IC50值达10.09 ng/ml,其与ZEN毒素呈线性相关(r=0.990);ZEN抗独特型单抗与莱克多巴胺、重金属铅、铬及虾过敏原单抗均无交叉反应,特异性良好。结论已成功制备玉米赤霉烯酮抗独特型单抗,该抗体与ZEN间存在"内影像"关系,可以替代ZEN毒素标准品,用于建立ZEN的无毒免疫学检测技术。     

4-甲基苯酚多克隆抗体的制备及应用

针对4-甲基苯酚(4-MP)的结构特点及半抗原设计原则,选择对羟基苯丙酸(34-HPA)作为半抗原,通过活化酯法与牛血清白蛋白(BSA)偶联合成人工免疫原,免疫3～4 m的雄性大白兔,制得多克隆抗体效价为1∶204 800。同时采用混合酸酐法合成半抗原-卵清蛋白(OVA)偶联物作为包被原,以4-MP为竞争的抗原,建立间接竞争ELISA检测方法。实验结果表明,理想工作条件为:包被原浓度1.05μg/mL,抗体稀释倍数1∶12 800,酶标二抗稀释倍数1∶1 000,0.1%明胶封闭1.5 h,竞争时间与酶标二抗反应时间均为1.0 h。4-甲基苯酚在δ1.0×10-2～1.0×103的范围内呈线性相关,得标准曲线方程y=-12.76x+66.66,最低检测限δ0.02,板内差异为7.0%,板间差异为8.4%。     

抗单端孢霉烯族T-2毒素单克隆抗体的制备及鉴定

通过将T-2毒素与戊二酸酐在吡啶中反应生成T-2HG中间体(TLC验证大部分的T-2毒素都已转化为T-2HG),然后采用活泼酯法将T-2HG中间体和载体蛋白BSA/OVA共价偶联,紫外扫描结果显示偶联成功。以合成的BSA偶联物(T-2HG-BSA)作为免疫原,免疫Balb/c小鼠,经细胞融合,采用改进的半固体培养基和两步筛选法共筛选到5株性能良好的单克隆细胞株,分别是4F12、2B5、2G7、1D7和4F3。其中2G7所分泌抗体的灵敏度(IC50)0.83μg/kg,分泌抗体的性能稳定,而且与ZEN、AFB1、OTA、DON均无交叉反应或交叉反应率很低。本研究为研发粮油产品中T-2毒素特异性免疫分析技术及产品奠定了重要基础。     

烟嘧磺隆人工抗原的合成与鼠源多克隆抗体的制备

为了制备烟嘧磺隆的多克隆抗体、建立烟嘧磺隆残留的免疫分析方法,以2-氨基磺酰基-N,N-二甲基烟酰胺(ASDM)和琥珀酸酐为原料合成半抗原,采用活泼酯法制备人工抗原并免疫小鼠。结果表明:经紫外可见吸收光谱扫描和聚丙烯酰胺凝胶电泳(SDS-PAGE)鉴定,人工抗原合成成功,半抗原与卵清蛋白(OVA)、牛血清白蛋白(BSA)的偶联比分别为10.5∶1和26.9∶1。5次免疫后小鼠抗血清的效价均达到1∶16 000以上,以ASDM-BSA为免疫原的小鼠产生了抑制烟嘧磺隆的多克隆抗体,建立的间接竞争ELISA方法对烟嘧磺隆的检测范围是1ng·mL-1~10μg·mL-1。为进一步建立更为高效、灵敏和方便的烟嘧磺隆ELISA检测方法提供了参考。     

Insect antifeedant properties of anthranoids from the genusVismia

Vismiones and ferruginins, representatives of a new class of lypophilic anthranoids from the genusVismia were found to inhibit feeding in larvae of species ofSpodoptera, Heliothis, and inLocusta migratoria.     

Direct observation of freeze-thaw instability of latex coatings via high pressure freezing and cryogenic SEM

Despite its industrial importance, the subject of freeze-thaw (F/T) stability of latex coatings has not been studied extensively. There is also a lack of fundamental understanding about the process and the mechanisms through which a coating becomes destabilized. High pressure (2100 bar) freezing fixes the state of water-suspended particles of polymer binder and inorganic pigments without the growth of ice crystals during freezing that produce artifacts in direct imaging scanning electron microscopy (SEM) of fracture surfaces of frozen coatings. We show that by incorporating copolymerizable functional monomers, it is possible to achieve F/T stability in polymer latexes and in low-VOC paints, as judged by the microstructures revealed by the cryogenic SEM technique. Particle coalescence as well as pigment segregation in F/T unstable systems are visualized. In order to achieve F/T stability in paints, latex particles must not flocculate and should provide protection to inorganic pigment and extender particles. Because of the unique capabilities of the cryogenic SEM, we are able to separate the effects of freezing and thawing, and study the influence of the rate of freezing and thawing on F/T stability. Destabilization can be caused by either freezing or thawing. A slow freezing process is more detrimental to F/T stability than a fast freezing process; the latter actually preserves suspension stability during freezing. Presented at the 82nd Annual Meeting of the Federation of Societies for Coatings Technology, October 27–29, 2004 in Chicago, IL. Tied for first place in The John A. Gordon Best Paper Competition.     

Ethanol and (−)-α-Pinene: Attractant Kairomones for Bark and Ambrosia Beetles in the Southeastern US

In 2002–2004, we examined the flight responses of 49 species of native and exotic bark and ambrosia beetles (Coleoptera: Scolytidae and Platypodidae) to traps baited with ethanol and/or (−)-α-pinene in the southeastern US. Eight field trials were conducted in mature pine stands in Alabama, Florida, Georgia, North Carolina, and South Carolina. Funnel traps baited with ethanol lures (release rate, about 0.6 g/day at 25–28°C) were attractive to ten species of ambrosia beetles (Ambrosiodmus tachygraphus, Anisandrus sayi, Dryoxylon onoharaensum, Monarthrum mali, Xyleborinus saxesenii, Xyleborus affinis, Xyleborus ferrugineus, Xylosandrus compactus, Xylosandrus crassiusculus, and Xylosandrus germanus) and two species of bark beetles (Cryptocarenus heveae and Hypothenemus sp.). Traps baited with (−)-α-pinene lures (release rate, 2–6 g/day at 25–28°C) were attractive to five bark beetle species (Dendroctonus terebrans, Hylastes porculus, Hylastes salebrosus, Hylastes tenuis, and Ips grandicollis) and one platypodid ambrosia beetle species (Myoplatypus flavicornis). Ethanol enhanced responses of some species (Xyleborus pubescens, H. porculus, H. salebrosus, H. tenuis, and Pityophthorus cariniceps) to traps baited with (−)-α-pinene in some locations. (−)-α-Pinene interrupted the response of some ambrosia beetle species to traps baited with ethanol, but only the response of D. onoharaensum was interrupted consistently at most locations. Of 23 species of ambrosia beetles captured in our field trials, nine were exotic and accounted for 70–97% of total catches of ambrosia beetles. Our results provide support for the continued use of separate traps baited with ethanol alone and ethanol with (−)-α-pinene to detect and monitor common bark and ambrosia beetles from the southeastern region of the US.     

Methyl 2,4,6-decatrienoates Attract Stink Bugs and Tachinid Parasitoids

Halyomorpha halys (Stål) (Pentatomidae), called the brown marmorated stink bug (BMSB), is a newly invasive species in the eastern USA that is rapidly spreading from the original point of establishment in Allentown, PA. In its native range, the BMSB is reportedly attracted to methyl (E,E,Z)-2,4,6-decatrienoate, the male-produced pheromone of another pentatomid common in eastern Asia, Plautia stali Scott. In North America, Thyanta spp. are the only pentatomids known to produce methyl 2,4,6-decatrienoate [the (E,Z,Z)-isomer] as part of their pheromones. Methyl 2,4,6-decatrienoates were field-tested in Maryland to monitor the spread of the BMSB and to explore the possibility that Thyanta spp. are an alternate host for parasitic tachinid flies that use stink bug pheromones as host-finding kairomones. Here we report the first captures of adult and nymph BMSBs in traps baited with methyl (E,E,Z)-2,4,6-decatrienoate in central Maryland and present data verifying that the tachinid, Euclytia flava (Townsend), exploits methyl (E,Z,Z)-2,4,6-decatrienoate as a kairomone. We also report the unexpected finding that various isomers of methyl 2,4,6-decatrienoate attract Acrosternum hilare (Say), although this bug apparently does not produce methyl decatrienoates. Other stink bugs and tachinids native to North America were also attracted to methyl 2,4,6-decatrienoates. These data indicate there are Heteroptera in North America in addition to Thyanta spp. that probably use methyl 2,4,6-decatrienoates as pheromones. The evidence that some pentatomids exploit the pheromones of other true bugs as kairomones to find food or to congregate as a passive defense against tachinid parasitism is discussed.     

2007～2008年世界塑料工业进展

收集了2007年7月～2008年6月世界塑料工业的相关资料,介绍了2007～2008年国外塑料工业的发展情况,提供了世界塑料产量、消费量及全球各类树脂的需求量及产能情况.按通用热塑性树脂(聚乙烯、聚丙烯、聚苯乙烯、聚氯乙烯、ABS树脂)、工程塑料(尼龙、聚碳酸酯、聚甲醛、热塑性聚酯、聚苯醚)、特种工程塑料(聚苯·硫醚、液晶聚合物、聚醚醚酮)、通用热固性树脂(酚醛、聚氨酯、不饱和聚酯树脂、环氧树脂)不同品种的顺序,对树脂的产量、消费量、供需状况及合成工艺、产品应用开发、树脂品种的延伸及应用的进一步扩展等技术作了详细介绍.     

2005～2006年国外塑料工业进展

收集了2005年7月～2006年6月国外塑料工业的相关资料,介绍了2005—2006年国外塑料工业的发展情况。提供了世界塑料产量、消费量及全球各类树脂生产量以及各国塑料制品的进出口情况。作为对比,介绍了中国塑料的生产情况。按通用热塑性树脂（聚乙烯、聚丙烯、聚苯乙烯、聚氯乙烯、ABS树脂）、工程塑料（聚酰胺、聚碳酸酯、聚甲醛、热塑性聚酯、聚苯醚）、通用热固性树脂（酚醛、聚氨酯、不饱和树脂、环氧树脂）、特种工程塑料（聚苯硫醚、液晶聚合物、聚醚醚酮）的品种顺序,对树脂的产量、消费量、供需状况及合成工艺、产品开发、树脂品种的延伸及应用的扩展作了详细的介绍。