排序方式: 共有35条查询结果,搜索用时 15 毫秒
1.
P. Galeffi G. Giunta S. Guida C. Cantale 《European journal of plant pathology / European Foundation for Plant Pathology》2002,108(5):479-483
Citrus tristeza virus (CTV) is one of the most destructive citrus virus diseases in the world. The construction of an engineered antibody, EMBL accession number AJ278109, able to specifically recognize its antigen, i.e. the coat protein of CTV, directly on infected plant material without any purification or manipulation of the entire woody plant. The potential uses of this engineered antibody are discussed. 相似文献
2.
Dezorzová-Tomanová K Molinková D Pekarová M Celer V Smola J 《Research in veterinary science》2007,83(1):85-90
Single-chain antibodies (scFv) exhibiting specific binding to Lawsonia intracellularis were isolated from a phagemid library expressing scFvs molecules on the surface of filamentous bacteriophages. For scFv selection whole bacterial cells were used and individual clones were tested in ELISA test. The total of seven unique clones with different fingerprint profiles was isolated. All clones were able to bind specifically in immunofluorescence assay. This is the first report of species specific recombinant antibodies against L. intracellularis. 相似文献
3.
Wouter van Wyngaardt Cordelia Mashau Isabel Wright Jeanni Fehrsen 《Journal of veterinary science (Suw?n-si, Korea)》2013,14(1):95-98
There is an ongoing need for standardized, easily renewable immunoreagents for detecting African horsesickness virus (AHSV). Two phage displayed single-chain variable fragment (scFv) antibodies, selected from a semi-synthetic chicken antibody library, were used to develop double antibody sandwich enzyme-linked immunosorbent assays (DAS-ELISAs) to detect AHSV. In the DAS-ELISAs, the scFv previously selected with directly immobilized AHSV-3 functioned as a serotype-specific reagent that recognized only AHSV-3. In contrast, the one selected with AHSV-8 captured by IgG against AHSV-3 recognized all nine AHSV serotypes but not the Bryanston strain of equine encephalosis virus. Serving as evidence for its serogroup-specificity. These two scFvs can help to rapidly confirm the presence of AHSV while additional serotype-specific scFvs may simplify AHSV serotyping. 相似文献
4.
Antibody phage display libraries are a useful tool in proteomic analyses. This study evaluated an antibody recombinant library for identification of sex-specific proteins on the sperm cell surface. The Griffin.1 library was used to produce phage antibodies capable of recognizing membrane proteins from Nelore sperm cells. After producing soluble monoclonal scFv, clones were screened on Simental sperm cells by flow cytometry and those that bound to 40–60% of cells were selected. These clones were re-analyzed using Nelore sperm cells and all clones bound to 40–60% of cells. Positive clones were submitted to a binding assay against male and female bovine leukocytes by flow cytometry and one clone preferentially bound to male cells. The results indicate that phage display antibodies are an alternative method for identification of molecules markers on sperm cells. 相似文献
5.
利用Bac-to-Bac杆状病毒表达系统,在Sf9昆虫细胞中表达抗对硫磷单链抗体,并评价该重组抗体scFv-4C6的分子识别活性。以分泌能特异性识别对硫磷的单克隆抗体的杂交瘤细胞株4C6为RNA来源,采用RT-PCR方法扩增抗体的重链和轻链可变区基因,经重叠延伸PCR方法串联拼接获得单链抗体基因片段(scFv-4C6)。构建包含目的片段的重组杆粒Bacmid-scFv-4C6,转染Sf9细胞表达目的蛋白,采用免疫印迹法(Western blotting)检测表达产物,间接竞争酶联免疫吸附(ic-ELISA)法评价产物的生物活性。结果表明:scFv-4C6基因片段拼装正确,成功转染Sf9细胞,并在转染后72 h表达量最高,表达的单链抗体大小为28.3 kD;表达产物能特异性识别对硫磷,IC50值为7.9 ng/mL,对甲基对硫磷和杀螟硫磷分别有12%和1.8%的交叉反应率,与亲本单克隆抗体的识别性能相似。该研究表明,具有生物活性的抗对硫磷单链抗体scFv-4C6可在昆虫细胞中成功得到表达。 相似文献
6.
单链抗体(Single chain variable fragment,scFv)是目前最受关注的基因重组抗体分子,是由重链可变区和轻链可变区以一个柔性肽段连接而成的最小抗体片段,它较好的保持着原代抗体的亲和特性,故而在农兽药残留检测方面具有潜在的巨大应用价值.文章综述了单链抗体技术、噬菌体展示和核糖体展示技术以及目前... 相似文献
7.
E. Sunderasan Rusni A. Kadir Valérie Pujade-Renaud Frédéric de Lamotte H. Y. Yeang Sheila Nathan 《Journal of General Plant Pathology》2009,75(1):19-26
Corynespora leaf disease poses a serious threat to rubber cultivation because infected leaves develop necrotic lesions and
abscise, leaving the tree unproductive. The destructiveness of Corynespora cassiicola has been largely attributed to cassiicolin, a protein toxin secreted by the fungus. Recombinant antibody technology offers
hope to curtail the disease whereby single-chain variable fragments (scFv) specific to cassiicolin could bind and deactivate
the toxin in genetically modified rubber trees that harbour the antibody gene. A scFv phage library was constructed from heavy
and light variable chains of IgG from cassiicolin immunized Balb/C mice spleen. Biopanning of the phage library yielded a
scFv clone with high specificity to cassiicolin. The nucleotide sequence and deduced amino acid sequence information of the
scFv were obtained. Hemagglutinin (HA)-tagged scFv expressed in Escherichia coli is discerned as a band at ca. 30 kDa on SDS-PAGE, and the corresponding band was detected by anti-HA IgG on a Western immunoblot.
Deactivation of cassiicolin by the affinity-purified scFv was demonstrated in a detached-leaf bio-assay on selected susceptible
Hevea clones (PB 260, RRIM 2020, RRIM 901 and RRIM 929). The assay was also performed on clones that are relatively more resistant
to the fungus (RRIM 600 and GT-1), and the results were as expected. Thus, we have successfully demonstrated that the cassicolin-specific
scFv can effectively reduce cassicolin toxicity. 相似文献
8.
从一个抗脂磷酸聚糖IgM型单克隆抗体细胞的mRNA中经RT-PCR克隆重链可变区(VH)和轻链可变区(VL),然后将VH、Linker和VL连接成scFv,并克隆到载体pCANTAB-5E,转Escherichia coliTG1感受态细胞,构建了单链抗体库。通过辅助噬菌体M13KO7感染后,使单链抗体展示在噬菌体衣壳蛋白pIII的N端,得到噬菌体展示的抗体库。将抗体库用于"淘洗—富集—扩增"3轮以后,得到针对脂磷酸聚糖特异性的噬菌体抗体。测序结果表明,该scFv的VH基因序列全长390 bp,编码127个氨基酸;VL基因序列全长341 bp,编码113个氨基酸。二者均符合小鼠免疫球蛋白可变区基因特征,含有4个框架区(FR)、3个抗原互补决定区(CDR)及抗体特征性的2个半胱氨酸残基。 相似文献
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10.
Bhaskaran S Jay CM Berghman LR Wagner GG Waghela SD 《Veterinary research communications》2005,29(6):463-476
Bovine colibacillosis caused by enterotoxigenic Escherichia coli (ETEC) is a worldwide problem. Adhesion of ETEC to intestinal cell receptors mediated by the surface protein F5 fimbriae
is the initial step in the establishment of colibacillosis. Prevention of ETEC F5+ adhesion to enterocytes protects newborn calves against collibacillosis. On the enterocytes, the F5 fimbriae bind to a ganglioside
that is also found on horse red blood cells. Thus, the presence of F5 fimbriae induces haemagglutination, which is useful
as an indicator in a functional assay system. In this study, recombinant anti-F5 scFv antibody fragment produced in E. coli HB2151 reacted with F5 fimbriae in ELISA and Western immunoblot, and prevented haemagglutination induced by the binding of
the F5 fimbriae to its natural host receptors on horse red blood cells. Given the ease with which recombinant antibodies can
be mass-produced, the presently described scFv may hold promise as a prophylactic agent for colibacillosis. 相似文献