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国家自然科学基金(81028010)

作品数:2 被引量:4H指数:1
相关作者:徐多多李思静吴靖潘鹏涛王丽更多>>
相关机构:东北师范大学更多>>
发文基金:国家自然科学基金长春市科技发展计划项目更多>>
相关领域:生物学农业科学更多>>

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重组人血管内皮生长因子(VEGF)单克隆抗体的制备被引量:1
2013年
为制备人血管内皮生长因子(VEGF)单克隆抗体,制备并纯化了VEGF蛋白,通过免疫、细胞融合、筛选等方法获得了能够稳定分泌单克隆抗体的细胞株;采用小鼠腹腔接种法制备腹水,并用G蛋白(Protein G)亲和层析柱对抗体进行了纯化;采用间接ELISA及Western blot实验分别对纯化后的抗体进行了效价测定和特异性验证.结果表明:此方法可成功筛选出能稳定分泌VEGF单克隆抗体的杂交瘤细胞株,纯化后腹水抗体效价为1∶1 024;Western blot检验证实制备的单克隆抗体能特异性识别VEGF蛋白.
徐多多李思静李芳芊潘鹏涛吴靖王丽
关键词:VEGF单克隆抗体ELISA
Nontoxic virus nanofibers improve the detection sensitivity for the anti-p53 antibody, a biomarker in cancer patients被引量:3
2015年
The presence of anti-p53 antibody in serum is a biomarker for cancer. However, its high sensitivity detection is still an issue in cancer diagnosis. To tackle this challenge, we used fd phage, a human-safe bacteria-specific virus nanofiber that can be mass-produced by infecting host bacteria in an error-free manner, and genetically engineered it to display a peptide capable of recognizing and capturing anti-p53 antibody on its side wall. We employed the resultant phage nanofibers as a capture probe to develop a modified version of the enzyme- linked immunosorbent assay (ELISA) method, termed phage-ELISA. We compared it to the traditional ELISA method for the detection of anti-p53 antibody, p53-ELISA, which uses recombinant wild-type p53 protein to capture anti-p53 antibody. We applied phage-ELISA to detect anti-p53 antibody in an experimental group of 316 patients with various types of malignant tumors. We found that a detection rate of 17.7% (56 positive cases) was achieved by phage-ELISA, which was comparable to the detection rate of 20.6% for p53-ELISA (65 positive cases). However, when both phage and p53 were combined to form antibody-capturing probes for phage/p53-ELISA, a detection rate of 30.4% (96 positive cases) was achieved. Our work showed that owing to the combined capture of the anti-p53 antibody by both phage nanofibers and p53, the phage/p53-ELISA achieved the highest diagnostic accuracy and detection efficiency for the anti-p53 antibody in patients with various types of cancers. Our work suggests that a combination of nanofibers and antigens, both of which capture antibody, could lead to increased detection sensitivity, which is useful for applications in the life sciences, clinical medicine, and environmental sciences.
Pengtao PanYicun WangYe ZhuXiang GaoZhigang JuPenghe QiuLi WangChuanbin Mao
关键词:PHAGEVIRUSPROTEINNANOFIBERS
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