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小鼠Metrnl单克隆抗体的制备及鉴定

曲艺,  李志勇,  缪朝玉

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曲艺, 李志勇, 缪朝玉. 小鼠Metrnl单克隆抗体的制备及鉴定[J]. 药学实践与服务, 2017, 35(2): 126-129,192. doi: 10.3969/j.issn.1006-0111.2017.02.007
引用本文: 曲艺, 李志勇, 缪朝玉. 小鼠Metrnl单克隆抗体的制备及鉴定[J]. 药学实践与服务, 2017, 35(2): 126-129,192. doi: 10.3969/j.issn.1006-0111.2017.02.007
shu
QU Yi, LI Zhiyong, MIAO Chaoyu. Preparation and characterization of monoclonal antibodies against mouse Metrnl[J]. Journal of Pharmaceutical Practice and Service, 2017, 35(2): 126-129,192. doi: 10.3969/j.issn.1006-0111.2017.02.007
Citation: QU Yi, LI Zhiyong, MIAO Chaoyu. Preparation and characterization of monoclonal antibodies against mouse Metrnl[J]. Journal of Pharmaceutical Practice and Service, 2017, 35(2): 126-129,192. doi: 10.3969/j.issn.1006-0111.2017.02.007
shu

小鼠Metrnl单克隆抗体的制备及鉴定

doi: 10.3969/j.issn.1006-0111.2017.02.007

  • 第二军医大学药学院药理学教研室, 上海 200433

基金项目: 国家自然科学基金（81130061，81202572，81373414）；上海市科学技术委员会科研计划项目（16JC1405100）

Preparation and characterization of monoclonal antibodies against mouse Metrnl

  • Department of Pharmacology, School of Pharmacy, Second Military Medical University, Shanghai 200433, China

  • 摘要: 目的 制备抗小鼠Metrnl单克隆抗体，并进行初步筛选鉴定。 方法 分别制备小鼠Metrnl多肽片段和全长蛋白作为免疫小鼠抗原，取免疫后小鼠脾细胞与SP2/0骨髓瘤细胞融合，筛选阳性杂交瘤细胞，并亚克隆获得稳定细胞株，制备腹水。用ELISA方法检测腹水抗体效价；用Western blot方法鉴定抗体。 结果 由14种多肽抗原制备的56株单克隆抗体中，未筛选出可用于Western blot识别Metrnl的抗体；由全长蛋白制备的25株抗体中，有12株可识别Metrnl蛋白。 结论 本实验成功制备了12株单抗，可用于识别检测小鼠Metrnl蛋白。
    Abstract: Objective To prepare monoclonal antibodies (mAb) against mouse Metrnl and identify its specificity. Methods Mouse Metrnl polypeptide fragments and full-length protein were prepared as antigens to immunize mice.Then mice spleen cells were fused with SP2/0 myeloma cells to obtain hybridoma cells which were screened for positive clone in order to subclone for stable cell lines.After ascites were prepared, ELISA method was used to detect the antibodies titer.Western blot method was applied to identify their specificity. Results No effective antibodies were identified from the ascites derived from 14 polypeptide antigens.Among the 25 antibodies derived from the full-length protein, 12 monoclonal antibodies can be used to identify the recombinant Metrnl protein. Conclusion 12 monoclonal antibodies were successfully prepared to identify mouse Metrnl protein.
  • [1] Li ZY,Zheng SL,Wang P,et al.Subfatin is a novel adipokine and unlike Meteorin in adipose and brain expression[J].CNS Neurosci Ther,2014,20(4):344-354.
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    [5] Jorgensen JR,Fransson A,Fjord-Larsen L,et al.Cometin is a novel neurotrophic factor that promotes neurite outgrowth and neuroblast migration in vitro and supports survival of spiral ganglion neurons in vivo[J].Exp Neurol,2012,233(1):172-181.
    [6] Rao RR,Long JZ,White JP,et al.Meteorin-like is a hormone that regulates immune-adipose interactions to increase beige fat thermogenesis[J].Cell,2014,157(6):1279-1291.
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    [9] 户义, 刘雪松,朱勇,等.抗Ig融合蛋白Fc段单克隆抗体的制备、鉴定与应用研究[J].细胞与分子免疫学杂志,2003,19(2):170-171,175.
    [10] 许晓光, 朱勇,刘雪松,等.抗人CD100单克隆抗体的制备与初步鉴定[J].细胞与分子免疫学杂志,2003,19(1):80-82.
    [11] 杜春红, 钟佑宏,陈平,等.BALB/c小鼠选择与腹水产量和抗体效价间关系的研究[J].实验动物科学,2008,25(6):9-11.
    [12] Li ZY,Fan MB,Zhang SL,et al.Intestinal Metrnl released into the gut lumen acts as a local regulator for gut antimicrobial peptides[J].Acta Pharmacol Sin,2016,37(11):1458-1466.
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  • 收稿日期:  2016-11-08
  • 修回日期:  2016-12-23

小鼠Metrnl单克隆抗体的制备及鉴定

doi: 10.3969/j.issn.1006-0111.2017.02.007
  • 第二军医大学药学院药理学教研室, 上海 200433
    • 基金项目:  国家自然科学基金（81130061，81202572，81373414）；上海市科学技术委员会科研计划项目（16JC1405100）
  • 收稿日期: 2016-11-08
  • 修回日期: 2016-12-23

摘要: 目的 制备抗小鼠Metrnl单克隆抗体，并进行初步筛选鉴定。 方法 分别制备小鼠Metrnl多肽片段和全长蛋白作为免疫小鼠抗原，取免疫后小鼠脾细胞与SP2/0骨髓瘤细胞融合，筛选阳性杂交瘤细胞，并亚克隆获得稳定细胞株，制备腹水。用ELISA方法检测腹水抗体效价；用Western blot方法鉴定抗体。 结果 由14种多肽抗原制备的56株单克隆抗体中，未筛选出可用于Western blot识别Metrnl的抗体；由全长蛋白制备的25株抗体中，有12株可识别Metrnl蛋白。 结论 本实验成功制备了12株单抗，可用于识别检测小鼠Metrnl蛋白。

English Abstract

曲艺, 李志勇, 缪朝玉. 小鼠Metrnl单克隆抗体的制备及鉴定[J]. 药学实践与服务, 2017, 35(2): 126-129,192. doi: 10.3969/j.issn.1006-0111.2017.02.007
引用本文: 曲艺, 李志勇, 缪朝玉. 小鼠Metrnl单克隆抗体的制备及鉴定[J]. 药学实践与服务, 2017, 35(2): 126-129,192. doi: 10.3969/j.issn.1006-0111.2017.02.007
shu
QU Yi, LI Zhiyong, MIAO Chaoyu. Preparation and characterization of monoclonal antibodies against mouse Metrnl[J]. Journal of Pharmaceutical Practice and Service, 2017, 35(2): 126-129,192. doi: 10.3969/j.issn.1006-0111.2017.02.007
Citation: QU Yi, LI Zhiyong, MIAO Chaoyu. Preparation and characterization of monoclonal antibodies against mouse Metrnl[J]. Journal of Pharmaceutical Practice and Service, 2017, 35(2): 126-129,192. doi: 10.3969/j.issn.1006-0111.2017.02.007
shu

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