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SHN3多克隆抗体纯化与鉴定
  • ISSN号:1004-311X
  • 期刊名称:《生物技术》
  • 时间:0
  • 分类:Q511[生物学—生物化学]
  • 作者机构:[1]上海大学生命科学学院,上海200444, [2]上海大学系统生物学技术研究所,上海200444
  • 相关基金:基金项目:国家自然科学基金项目(“Dcf1基因对大脑发育神经元迁移及树突棘形成机制的研究”,No.81271253;“Dcf1基因在大脑发育过程的作用机制”,No.31070954)资助
中文摘要:

目的:利用SHN3免疫特异肽段亲和层析纯化抗SHN3多克隆抗体。方法:将制备多克隆抗体SHN3免疫原片段(99bp)划分为4段互相重叠等长的基因片段(48bp),构建原核表达重组质粒并转化E.coil的B121(DE3)菌株。IPGT诱导重组蛋白表达,Western blotting筛选并人工合成与抗体免疫结合特异性片段,亲和层析纯化抗体,Western blotting检测纯化结果。结果:成功地筛选到SHN3免疫原特异肽段part-3,以亲和层析的方式纯化多克隆抗体,纯化后免疫特异性提高。结论:亲和层析的方式纯化得到的SHN3多克隆抗体抗体,有效地提高免疫特异性,为后续SHN3蛋白功能研究和应用奠定了基础。

英文摘要:

Objective:To purify polyclonal antibodies against SHN3 by affinity chromatography with immunologic specificity peptide frag- ment. Method:SHN3 immunologic specificity gene (90bp)was divided 4 equal length parts(48bp). And subcloned into prokaryotic ex- pression vector Pmal - C2. These recombinant vectors were transformed into E. coliBL21 ( DE3 ). Recombinant peptide expression were in- duced by IPTG. Western blotting method was used to screen the best immunologic specificity peptide fragment. The polyclonal antibodies was purified by affinity chromatography and checked by Western blotting. Result:The immunologic specificity peptide fragment part -3 of SHN3 was screened successfully, and the SHN3 polyclonal antibodies were purified with this peptide by affinity chromatography. The affini- ty chromatography of SHN3 polyclonal antibodies was improved significantly. Conclusion: Affinity chromatograph for SHN3 polyclonal anti- bodies purification was successfully established and immunologic specificity was significantly improved. The method laid a foundation for subsequent SHN3 research and application.

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期刊信息
  • 《生物技术》
  • 北大核心期刊(2014版)
  • 主管单位:黑龙江省科学院
  • 主办单位:黑龙江省科学院微生物研究所 黑龙江省微生物学会 黑龙江省生物工程学会
  • 主编:张介驰
  • 地址:哈尔滨市道里区兆麟街68号
  • 邮编:150010
  • 邮箱:swjszz@163.com
  • 电话:0451-84615121
  • 国际标准刊号:ISSN:1004-311X
  • 国内统一刊号:ISSN:23-1319/Q
  • 邮发代号:14-225
  • 获奖情况:
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  • 被引量:13503