小鼠Metrnl单克隆抗体的制备及鉴定

曲艺; 李志勇; 缪朝玉

doi:10.3969/j.issn.1006-0111.2017.02.007

留言板

尊敬的读者、作者、审稿人, 关于本刊的投稿、审稿、编辑和出版的任何问题, 您可以本页添加留言。我们将尽快给您答复。谢谢您的支持!

姓名
邮箱
手机号码
标题
留言内容
验证码

x 关闭永久关闭

应中央军委要求，2022年9月起，《药学实践杂志》将更名为《药学实践与服务》，双月刊，正文96页；2023年1月起，拟出版月刊，正文64页，数据库收录情况与原《药学实践杂志》相同。欢迎作者踊跃投稿！

小鼠Metrnl单克隆抗体的制备及鉴定

曲艺, 李志勇, 缪朝玉

文章导航 > 药学实践与服务 > 2017 > 35(2): 126-129,192

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

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

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。
- Metrnl /
- 杂交瘤技术 /
- 单克隆抗体
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.
- Metrnl /
- hybridization technique /
- monoclonal antibody

[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.
[2]	Nishino J,Yamashita K,Hashiguchi H,et al.Meteorin:a secreted protein that regulates glial cell differentiation and promotes axonal extension[J].EMBO J,2004,23(9):1998-2008.
[3]	Zheng SL,Li ZY,Song J,et al.Metrnl:a secreted protein with new emerging functions[J].Acta Pharmacol Sin,2016,37(5):571-579.
[4]	Li ZY,Song J,Zheng SL,et al.Adipocyte Metrnl antagonizes insulin resistance through PPAR gamma signaling[J].Diabetes,2015,64(12):4011-4022.
[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.
[7]	Gong W,Liu Y,Wu Z,et al.Meteorin-like shows unique expression pattern in bone and its overexpression inhibits osteoblast differentiation[J].PLoS One,2016,11(10):e0164446.
[8]	Deng A,Tan W,He S,et al.Monoclonal antibody-based enzyme linked immunosorbent assay for the analysis of jasmonates in plants[J].J Integr Plant Biol,2008,50(8):1046-1052.
[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.
[13]	Ushach I,Burkhardt AM,Martinez C,et al.METEORIN-LIKE is a cytokine associated with barrier tissues and alternatively activated macrophages[J].Clin Immunol,2015,156(2):119-127.
[14]	Yamada T.Therapeutic monoclonal antibodies[J].Keio J Med,2011,60(2):37-46.

[1]	徐飞, 陈瑾, 李志勇. 胰岛素增敏剂罗格列酮升高血液中镍纹样蛋白水平 . 药学实践与服务, 2024, 42(4): 165-168. doi: 10.12206/j.issn.2097-2024.202308053
[2]	王雪莲, 郑斯莉, 李志勇, 罗亨宇, 缪朝玉. 全身过表达人METRNL基因小鼠模型的构建与验证 . 药学实践与服务, 2024, 42(): 1-6. doi: 10.12206/j.issn.2097-2024.202311014
[3]	张馨月, 台宗光, 朱全刚, 陈中建, 鲍蕾蕾. 治疗黑色素瘤的DC疫苗研究新进展 . 药学实践与服务, 2023, 41(11): 643-647. doi: 10.12206/j.issn.2097-2024.202211043
[4]	夏晶, 陈行, 陈建明, 武鑫. 冰片辅助胶质瘤治疗的临床前研究进展 . 药学实践与服务, 2023, 41(4): 202-206. doi: 10.12206/j.issn.2097-2024.202203126
[5]	陈思敏, 凌期盛, 张赛龙, 缪朝玉. 肠上皮Metrnl基因敲除对肠道菌群以及溃疡性结肠炎的影响 . 药学实践与服务, 2023, 41(11): 654-661. doi: 10.12206/j.issn.2097-2024.202211050
[6]	马明华, 汪晓河, 戴媛媛, 顾小燕, 吴铁军, 年华. 夏枯草消瘤合剂对Lewis肺癌小鼠的药效作用 . 药学实践与服务, 2020, 38(1): 57-62. doi: 10.3969/j.issn.1006-0111.201812070
[7]	赵全凤, 龚玉竹, 谢赣丰, 夏培元, 唐敏. 临床药师参与晚期直肠神经内分泌瘤患者的药学服务 . 药学实践与服务, 2019, 37(6): 559-562. doi: 10.3969/j.issn.1006-0111.2019.06.017
[8]	夏涌, 寇晓霞, 杨喜晶, 周福平, 张迁, 钱其军. 程序死亡受体1抗体治疗18例晚期肿瘤患者的临床疗效观察及文献复习 . 药学实践与服务, 2017, 35(2): 174-177. doi: 10.3969/j.issn.1006-0111.2017.02.020
[9]	曲艺, 李志勇, 缪朝玉. 抗体的发展过程及临床应用 . 药学实践与服务, 2017, 35(4): 301-303,333. doi: 10.3969/j.issn.1006-0111.2017.04.004
[10]	郑洁, 朱珍珍, 虞如芬. rhGM-CSF凝胶对LEEP治疗宫颈上皮内瘤变创面愈合的影响 . 药学实践与服务, 2014, 32(2): 124-127. doi: 10.3969/j.issn.1006-0111.2014.02.013
[11]	谭何新, 刘颖, 张磊. 丹参果糖磷酸酶基因的克隆和功能研究 . 药学实践与服务, 2014, 32(1): 35-41,48. doi: 10.3969/j.issn.1006-0111.2014.01.009
[12]	刘莹, 厉建中, 张俊平. 人的核糖体蛋白S3a基因克隆、表达、纯化与亚细胞定位 . 药学实践与服务, 2011, 29(2): 97-100.
[13]	林观样, 郭红, 余磊. 红细胞在不同pH下致敏对头孢他啶针剂的IgM和IgG抗体检测结果的影响 . 药学实践与服务, 2010, 28(3): 206-208.
[14]	苏开仲, 马鸿媚, 魏文树. 复方鲨鱼软骨素对小鼠移植性肉瘤S₁₈₀抑瘤作用的拆方研究 . 药学实践与服务, 2007, (2): 80-81.
[15]	周全, 马光大, 管斐. 纳米技术与药物新剂型 . 药学实践与服务, 2003, (3): 140-142.
[16]	陈兴雅, 郭良耀, 赵小亭. 国产白介素-2并COHP方案联合化疗治疗恶性淋巴瘤的临床疗效观察 . 药学实践与服务, 1997, (4): 201-202.
[17]	李文茂. 医院输液生产的净化技术 . 药学实践与服务, 1995, (6): 375-376.
[18]	龚江宁, 董可因. 血细胞回输-治疗多发性骨髓瘤可能成功 . 药学实践与服务, 1995, (1): 7-7.
[19]	孙华君, 张紫洞. 如何避免艾滋病及抗体检查结果阳性怎么办 . 药学实践与服务, 1990, (3): 19-20.
[20]	叶凤山, 张紫洞. 单克隆抗体的抗移植排斥作用 . 药学实践与服务, 1989, (3): 60-60.

点击查看大图

计量

文章访问数: 2716
HTML全文浏览量: 240
PDF下载量: 359
被引次数: 0

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

关键词:

Metrnl /

杂交瘤技术 /

单克隆抗体

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。