抗体验证指南 2016

2016-09-09 Nat Methods. 2016 Sep 5. doi: 10.1038/nmeth.3995

据估计,因抗体质量不佳,生命科学领域每年的损失达到8亿美元。同时,这也造成了无数的实验失败,浪费了宝贵的样本和科学家的青春。尽管科学界已经意识到这个问题,但目前仍然缺乏抗体验证的整体框架。 为了刺激研究界实现更高标准的抗体重复性,国际抗体验证工作组(IWGAV)近日出台了一套指南。这些指南发表在9月5日的《Nature Methods》杂志上,强调了抗体验证需要针对应用和背景来开展。 这篇题为

中文标题:

抗体验证指南 2016

英文标题:

A proposal for validation of antibodies

发布机构:

发布日期:

2016-09-09

简要介绍:

据估计,因抗体质量不佳,生命科学领域每年的损失达到8亿美元。同时,这也造成了无数的实验失败,浪费了宝贵的样本和科学家的青春。尽管科学界已经意识到这个问题,但目前仍然缺乏抗体验证的整体框架。

为了刺激研究界实现更高标准的抗体重复性,国际抗体验证工作组(IWGAV)近日出台了一套指南。这些指南发表在9月5日的《Nature Methods》杂志上,强调了抗体验证需要针对应用和背景来开展。

这篇题为“A Proposal for Validation of Antibodies”的文章是由IWGAV的数名成员发表的,包括工作组主席、瑞典皇家理工学院的微生物学教授Mathias Uhlén。文章提出了五点,以指导特定应用中的抗体验证。

•遗传策略:测定对照细胞或组织中的相关信号,这些对照中的目标基因已利用CRISPR或RNAi等技术敲除或敲低。

•正交策略:利用不依赖抗体的方法定量多个样品,然后检查依赖抗体和不依赖抗体这两种定量之间的相关性。

•独立抗体策略:利用两个或更多的独立抗体来识别目标蛋白上的不同表位,再通过比较和定量分析来确认特异性。

•表达标记蛋白:修饰内源的目标基因,增加亲和标签或荧光蛋白的序列。标记蛋白的信号可与基于抗体的检测相关联。

•免疫捕获加上质谱分析:将免疫捕获与MS分析相结合,以确定与纯化的抗体直接作用的蛋白质,以及可能与目标蛋白形成复合物的蛋白质。

这篇文章还对生产商和使用者提出建议,以确保长期的抗体重复性。作者们建议使用者在自己特定的应用或样品中,至少运用上面的一种策略来开展验证。同时,他们还建议使用者按照一定的形式报告,包含货号、批号和RRID(研究资源标识符),以确保研究中使用的任何抗体可被明确识别。

同时,作者们也向生产商提出了类似的建议,至少运用一种策略来验证,并提供尽可能多的抗体信息,如鉴定检测中的抗体浓度、细胞提取物的浓度、抗体生产所用的免疫原、抗体-表位的亲和力、缓冲液配方等。

“这篇文章迈出了重要的第一步,有望制定出广泛接受的抗体验证标准,确保生命医学研究使用高质量且一致的抗体,”Uhlén博士表示。“我们期待抗体用户、出版商、资助机构和生产商的反馈,以帮助加强这个初步建议,并确保这些基本工具的可靠性。”

原始出处:

Uhlen M, Bandrowski A, Carr S, Edwards A, Ellenberg J, Lundberg E, Rimm DL, Rodriguez H, Hiltke T, Snyder M, Yamamoto T. A proposal for validation of antibodies. Nat Methods. 2016 Sep 5. doi: 10.1038/nmeth.3995

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[AttachmentFileName(sort=100, fileName=抗体验证指南 2016)] GetToolGuiderByIdResponse(projectId=1, id=b640e1c0012e82a7, title=抗体验证指南 2016, enTitle=A proposal for validation of antibodies, guiderFrom=Nat Methods. 2016 Sep 5. doi: 10.1038/nmeth.3995, authorId=null, author=, summary=据估计,因抗体质量不佳,生命科学领域每年的损失达到8亿美元。同时,这也造成了无数的实验失败,浪费了宝贵的样本和科学家的青春。尽管科学界已经意识到这个问题,但目前仍然缺乏抗体验证的整体框架。 为了刺激研究界实现更高标准的抗体重复性,国际抗体验证工作组(IWGAV)近日出台了一套指南。这些指南发表在9月5日的《Nature Methods》杂志上,强调了抗体验证需要针对应用和背景来开展。 这篇题为, cover=, journalId=null, articlesId=null, associationId=null, associationName=, associationIntro=, copyright=0, guiderPublishedTime=Fri Sep 09 20:24:01 CST 2016, originalUrl=, linkOutUrl=, content=据估计,因抗体质量不佳,生命科学领域每年的损失达到8亿美元。同时,这也造成了无数的实验失败,浪费了宝贵的样本和科学家的青春。尽管科学界已经意识到这个问题,但目前仍然缺乏抗体验证的整体框架。 <p>为了刺激研究界实现更高标准的抗体重复性,国际抗体验证工作组(IWGAV)近日出台了一套指南。这些指南发表在9月5日的《Nature Methods》杂志上,强调了抗体验证需要针对应用和背景来开展。</p> <p>这篇题为“A Proposal for Validation of Antibodies”的文章是由IWGAV的数名成员发表的,包括工作组主席、瑞典皇家理工学院的微生物学教授Mathias Uhlén。<span style="font-weight: bold;">文章提出了五点,以指导特定应用中的抗体验证。</span></p> <p><span style="font-weight: bold;">•遗传策略:</span>测定对照细胞或组织中的相关信号,这些对照中的目标基因已利用CRISPR或RNAi等技术敲除或敲低。</p> <p><span style="font-weight: bold;"> •正交策略:</span>利用不依赖抗体的方法定量多个样品,然后检查依赖抗体和不依赖抗体这两种定量之间的相关性。</p> <p><span style="font-weight: bold;"> •独立抗体策略:</span>利用两个或更多的独立抗体来识别目标蛋白上的不同表位,再通过比较和定量分析来确认特异性。</p> <p><span style="font-weight: bold;"> •表达标记蛋白:</span>修饰内源的目标基因,增加亲和标签或荧光蛋白的序列。标记蛋白的信号可与基于抗体的检测相关联。</p> <p><span style="font-weight: bold;"> •免疫捕获加上质谱分析:</span>将免疫捕获与MS分析相结合,以确定与纯化的抗体直接作用的蛋白质,以及可能与目标蛋白形成复合物的蛋白质。</p> <p>这篇文章还对生产商和使用者提出建议,以确保长期的抗体重复性。作者们建议使用者在自己特定的应用或样品中,至少运用上面的一种策略来开展验证。同时,他们还建议使用者按照一定的形式报告,包含货号、批号和RRID(研究资源标识符),以确保研究中使用的任何抗体可被明确识别。</p> <p>同时,作者们也向生产商提出了类似的建议,至少运用一种策略来验证,并提供尽可能多的抗体信息,如鉴定检测中的抗体浓度、细胞提取物的浓度、抗体生产所用的免疫原、抗体-表位的亲和力、缓冲液配方等。</p> <p>“这篇文章迈出了重要的第一步,有望制定出广泛接受的抗体验证标准,确保生命医学研究使用高质量且一致的抗体,”Uhlén博士表示。“我们期待抗体用户、出版商、资助机构和生产商的反馈,以帮助加强这个初步建议,并确保这些基本工具的可靠性。”</p> <p><span style="font-weight: bold;">原始出处:</span></p> <p>Uhlen M, Bandrowski A, Carr S, Edwards A, Ellenberg J, Lundberg E, Rimm DL, Rodriguez H, Hiltke T, Snyder M, Yamamoto T. <a href="http://www.ncbi.nlm.nih.gov/pubmed/27595404" ref="ordinalpos=1&ncbi_uid=27595404&link_uid=27595404&linksrc=docsum_title">A proposal for validation of antibodies.</a> <span class="jrnl" title="Nature methods">Nat Methods</span>. 2016 Sep 5. doi: 10.1038/nmeth.3995</p>, tagList=[TagDto(tagId=63, tagName=指南), TagDto(tagId=2445, tagName=抗体)], categoryList=[CategoryDto(categoryId=42, categoryName=转化医学, tenant=100), CategoryDto(categoryId=21100, categoryName=达仁堂循证e学界, tenant=100)], articleKeywordId=null, articleKeyword=null, articleKeywordNum=null, guiderKeywordId=null, guiderKeyword=null, guiderKeywordNum=null, haveAttachments=1, attachmentList=null, guiderType=0, guiderArea=指南, guiderLanguage=0, guiderRegion=1, opened=0, paymentType=null, paymentAmount=10, recommend=0, recommendEndTime=null, sticky=0, stickyEndTime=null, allHits=21905, appHits=82, showAppHits=0, pcHits=7902, showPcHits=14933, likes=186, shares=0, comments=5, approvalStatus=1, publishedTime=Fri Sep 09 20:24:01 CST 2016, publishedTimeString=2016-09-09, pcVisible=1, appVisible=1, editorId=4754896, editor=fbzhang, waterMark=0, formatted=0, memberCards=[], isPrivilege=0, deleted=0, version=1, createdBy=null, createdName=fbzhang, createdTime=Fri Sep 09 20:24:01 CST 2016, updatedBy=null, updatedName=null, updatedTime=Sat Jan 06 07:20:09 CST 2024, courseDetails=[], otherVersionGuiders=[], isGuiderMember=false, ipAttribution=, attachmentFileNameList=[AttachmentFileName(sort=100, fileName=抗体验证指南 2016)])
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    2016-09-11 天仙居

    有用

    0

  2. [GetPortalCommentsPageByObjectIdResponse(id=123162, encodeId=336b123162bf, content=有用, beContent=null, objectType=guider, channel=null, level=null, likeNumber=115, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=88ff1949572, createdName=天仙居, createdTime=Sun Sep 11 23:40:00 CST 2016, time=2016-09-11, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=123134, encodeId=9beb123134cc, content=很好,不错,以后会多学习, beContent=null, objectType=guider, channel=null, level=null, likeNumber=102, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/jW482SpianMayicTRbRZ5RzUn81b5CF5ibVPib8jWI92iciaxmqGHlwruOnK4SVZykd4iahqo1hicklIibyZoxm55X7EM8BW9eX1h1hQ2/0, createdBy=8aa81937526, createdName=李东泽, createdTime=Sun Sep 11 21:48:00 CST 2016, time=2016-09-11, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=122451, encodeId=175712245188, content=好,有用,下载了, beContent=null, objectType=guider, channel=null, level=null, likeNumber=1, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=http://cacheapi.medsci.cn/resource/upload/20160601/IMG574E13FEE55F33222.jpg, createdBy=929c1478678, createdName=zibozhouping, createdTime=Sun Sep 11 11:18:00 CST 2016, time=2016-09-11, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=121933, encodeId=b3da1219337d, content=继续学习, beContent=null, objectType=guider, channel=null, level=null, likeNumber=114, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/jW482SpianMayicTRbRZ5RzfPkmqRTH5rHib5mcib8Wb3QdXjhfOK1UubQL2lictjJFdFJ6Lc3cnnCOSWqxXZ4Kk6y5GdzDoy764R/0, createdBy=156e1644785, createdName=doctorJiangchao, createdTime=Sun Sep 11 06:52:00 CST 2016, time=2016-09-11, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=121934, encodeId=26b01219343f, content=继续关注, beContent=null, objectType=guider, channel=null, level=null, likeNumber=92, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/jW482SpianMayicTRbRZ5RzfPkmqRTH5rHib5mcib8Wb3QdXjhfOK1UubQL2lictjJFdFJ6Lc3cnnCOSWqxXZ4Kk6y5GdzDoy764R/0, createdBy=156e1644785, createdName=doctorJiangchao, createdTime=Sun Sep 11 06:52:00 CST 2016, time=2016-09-11, status=1, ipAttribution=)]
    2016-09-11 李东泽

    很好,不错,以后会多学习

    0

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    2016-09-11 zibozhouping

    好,有用,下载了

    0

  4. [GetPortalCommentsPageByObjectIdResponse(id=123162, encodeId=336b123162bf, content=有用, beContent=null, objectType=guider, channel=null, level=null, likeNumber=115, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=88ff1949572, createdName=天仙居, createdTime=Sun Sep 11 23:40:00 CST 2016, time=2016-09-11, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=123134, encodeId=9beb123134cc, content=很好,不错,以后会多学习, beContent=null, objectType=guider, channel=null, level=null, likeNumber=102, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/jW482SpianMayicTRbRZ5RzUn81b5CF5ibVPib8jWI92iciaxmqGHlwruOnK4SVZykd4iahqo1hicklIibyZoxm55X7EM8BW9eX1h1hQ2/0, createdBy=8aa81937526, createdName=李东泽, createdTime=Sun Sep 11 21:48:00 CST 2016, time=2016-09-11, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=122451, encodeId=175712245188, content=好,有用,下载了, beContent=null, objectType=guider, channel=null, level=null, likeNumber=1, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=http://cacheapi.medsci.cn/resource/upload/20160601/IMG574E13FEE55F33222.jpg, createdBy=929c1478678, createdName=zibozhouping, createdTime=Sun Sep 11 11:18:00 CST 2016, time=2016-09-11, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=121933, encodeId=b3da1219337d, content=继续学习, beContent=null, objectType=guider, channel=null, level=null, likeNumber=114, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/jW482SpianMayicTRbRZ5RzfPkmqRTH5rHib5mcib8Wb3QdXjhfOK1UubQL2lictjJFdFJ6Lc3cnnCOSWqxXZ4Kk6y5GdzDoy764R/0, createdBy=156e1644785, createdName=doctorJiangchao, createdTime=Sun Sep 11 06:52:00 CST 2016, time=2016-09-11, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=121934, encodeId=26b01219343f, content=继续关注, beContent=null, objectType=guider, channel=null, level=null, likeNumber=92, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/jW482SpianMayicTRbRZ5RzfPkmqRTH5rHib5mcib8Wb3QdXjhfOK1UubQL2lictjJFdFJ6Lc3cnnCOSWqxXZ4Kk6y5GdzDoy764R/0, createdBy=156e1644785, createdName=doctorJiangchao, createdTime=Sun Sep 11 06:52:00 CST 2016, time=2016-09-11, status=1, ipAttribution=)]
    2016-09-11 doctorJiangchao

    继续学习

    0

  5. [GetPortalCommentsPageByObjectIdResponse(id=123162, encodeId=336b123162bf, content=有用, beContent=null, objectType=guider, channel=null, level=null, likeNumber=115, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=null, createdBy=88ff1949572, createdName=天仙居, createdTime=Sun Sep 11 23:40:00 CST 2016, time=2016-09-11, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=123134, encodeId=9beb123134cc, content=很好,不错,以后会多学习, beContent=null, objectType=guider, channel=null, level=null, likeNumber=102, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/jW482SpianMayicTRbRZ5RzUn81b5CF5ibVPib8jWI92iciaxmqGHlwruOnK4SVZykd4iahqo1hicklIibyZoxm55X7EM8BW9eX1h1hQ2/0, createdBy=8aa81937526, createdName=李东泽, createdTime=Sun Sep 11 21:48:00 CST 2016, time=2016-09-11, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=122451, encodeId=175712245188, content=好,有用,下载了, beContent=null, objectType=guider, channel=null, level=null, likeNumber=1, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=http://cacheapi.medsci.cn/resource/upload/20160601/IMG574E13FEE55F33222.jpg, createdBy=929c1478678, createdName=zibozhouping, createdTime=Sun Sep 11 11:18:00 CST 2016, time=2016-09-11, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=121933, encodeId=b3da1219337d, content=继续学习, beContent=null, objectType=guider, channel=null, level=null, likeNumber=114, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/jW482SpianMayicTRbRZ5RzfPkmqRTH5rHib5mcib8Wb3QdXjhfOK1UubQL2lictjJFdFJ6Lc3cnnCOSWqxXZ4Kk6y5GdzDoy764R/0, createdBy=156e1644785, createdName=doctorJiangchao, createdTime=Sun Sep 11 06:52:00 CST 2016, time=2016-09-11, status=1, ipAttribution=), GetPortalCommentsPageByObjectIdResponse(id=121934, encodeId=26b01219343f, content=继续关注, beContent=null, objectType=guider, channel=null, level=null, likeNumber=92, replyNumber=0, topicName=null, topicId=null, topicList=[], attachment=null, authenticateStatus=null, createdAvatar=https://wx.qlogo.cn/mmopen/jW482SpianMayicTRbRZ5RzfPkmqRTH5rHib5mcib8Wb3QdXjhfOK1UubQL2lictjJFdFJ6Lc3cnnCOSWqxXZ4Kk6y5GdzDoy764R/0, createdBy=156e1644785, createdName=doctorJiangchao, createdTime=Sun Sep 11 06:52:00 CST 2016, time=2016-09-11, status=1, ipAttribution=)]
    2016-09-11 doctorJiangchao

    继续关注

    0