Enzyme-linked immunosorbent assay of changes in serum levels of growth hormone (cGH) in common carps (Cyprinus carpio)
文献类型：期刊论文
| 作者 | Wu, Gang1; Chen, LiHua2; Zhong, Shan1,3; Li, Qi2; Song, ChaoJun2; Jin, BoQuan2; Zhu, ZuoYan1 |
| 刊名 | SCIENCE IN CHINA SERIES C-LIFE SCIENCES
 |
| 出版日期 | 2008-02-01 |
| 卷号 | 51期号:2页码:157-163 |
| 关键词 | common carp growth hormone monoclonal antibodies ELISA starvation net cage |
| ISSN号 | 1006-9305 |
| 通讯作者 | Zhu, ZY, Wuhan Univ, Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China |
| 中文摘要 | The aim of the present study was to purify the common native carp growth hormone (ncGH), produce monoclonal antibodies (mAbs) to common native carp growth hormone (ncGH), and further enhance the sensitivity of enzyme-linked immunosorbent assays (ELISA) for ncGH. Additionally, we investigated changes in serum ncGH levels in carps raised in different environmental conditions. The recombinant grass carp (Ctenopharyngodon idella) growth hormone was purified and used as antigen to immunize the rabbit. The natural ncGH was isolated from the pituitaries of common carp. SDS-PAGE and Western blot utilizing the polyclonal anti-rgcGH antibody confirmed the purification of ncGH from pituitaries. Purified ncGH was then used as an immunogen in the B lymphocyte hybridoma technique. A total of 14 hybridoma cell lines (FMU-cGH 1-14) were established that were able to stably secrete mAbs against ncGH. Among them, eight clones (FMU-cGH1-6, 12 and 13) were successfully used for Western blot while nine clones (FMU-cGH 1-7, 9 and 10) were used in fluorescent staining and immunohistochemistry. Epitope mapping by competitive ELISA demonstrated that these mAbs recognized five different epitopes. A sensitive sandwich ELISA for detection of ncGH was developed using FMU-cGH12 as the coating mAb and FMU-cGH6 as the enzyme labeled mAb. This detection system was found to be highly stable and sensitive, with detection levels of 70 pg/mL. Additionally, we found that serum ncGH levels in restricted food group and in the net cage group increased 6.9-and 5.8-fold, respectively, when compared to controls, demonstrating differences in the GH stress response in common carp under different living conditions. |
| 英文摘要 | The aim of the present study was to purify the common native carp growth hormone (ncGH), produce monoclonal antibodies (mAbs) to common native carp growth hormone (ncGH), and further enhance the sensitivity of enzyme-linked immunosorbent assays (ELISA) for ncGH. Additionally, we investigated changes in serum ncGH levels in carps raised in different environmental conditions. The recombinant grass carp (Ctenopharyngodon idella) growth hormone was purified and used as antigen to immunize the rabbit. The natural ncGH was isolated from the pituitaries of common carp. SDS-PAGE and Western blot utilizing the polyclonal anti-rgcGH antibody confirmed the purification of ncGH from pituitaries. Purified ncGH was then used as an immunogen in the B lymphocyte hybridoma technique. A total of 14 hybridoma cell lines (FMU-cGH 1-14) were established that were able to stably secrete mAbs against ncGH. Among them, eight clones (FMU-cGH1-6, 12 and 13) were successfully used for Western blot while nine clones (FMU-cGH 1-7, 9 and 10) were used in fluorescent staining and immunohistochemistry. Epitope mapping by competitive ELISA demonstrated that these mAbs recognized five different epitopes. A sensitive sandwich ELISA for detection of ncGH was developed using FMU-cGH12 as the coating mAb and FMU-cGH6 as the enzyme labeled mAb. This detection system was found to be highly stable and sensitive, with detection levels of 70 pg/mL. Additionally, we found that serum ncGH levels in restricted food group and in the net cage group increased 6.9-and 5.8-fold, respectively, when compared to controls, demonstrating differences in the GH stress response in common carp under different living conditions. |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
| 学科主题 | Biology |
| 类目[WOS] | Biology |
| 研究领域[WOS] | Life Sciences & Biomedicine - Other Topics |
| 关键词[WOS] | CONFINEMENT STRESS ; RADIOIMMUNOASSAY ; VALIDATION ; RECEPTOR ; FISH ; PITUITARY ; LIVER |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000252876000009 |
| 公开日期 | 2010-10-13 |
| 源URL | [http://ir.ihb.ac.cn/handle/152342/8284]  |
| 专题 | 水生生物研究所_中科院水生所知识产出（2009年前）_期刊论文 |
| 作者单位 | 1.Wuhan Univ, Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol & Biotechnol, Wuhan 430072, Peoples R China 2.Fourth Mil Med Univ, Dept Immunol, Xian 710032, Peoples R China 3.Wuhan Univ, Coll Life Sci, Wuhan 430072, Peoples R China |
| 推荐引用方式 GB/T 7714 | Wu, Gang,Chen, LiHua,Zhong, Shan,et al. Enzyme-linked immunosorbent assay of changes in serum levels of growth hormone (cGH) in common carps (Cyprinus carpio)[J]. SCIENCE IN CHINA SERIES C-LIFE SCIENCES,2008,51(2):157-163. |
| APA | Wu, Gang.,Chen, LiHua.,Zhong, Shan.,Li, Qi.,Song, ChaoJun.,...&Zhu, ZuoYan.(2008).Enzyme-linked immunosorbent assay of changes in serum levels of growth hormone (cGH) in common carps (Cyprinus carpio).SCIENCE IN CHINA SERIES C-LIFE SCIENCES,51(2),157-163. |
| MLA | Wu, Gang,et al."Enzyme-linked immunosorbent assay of changes in serum levels of growth hormone (cGH) in common carps (Cyprinus carpio)".SCIENCE IN CHINA SERIES C-LIFE SCIENCES 51.2(2008):157-163. |
入库方式： OAI收割
来源：水生生物研究所
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