Development of a specific ELISA to measure BACE1 levels in human tissues.
Development of a specific ELISA to measure BACE1 levels in human tissues.
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DOI：
10.1016/j.jneumeth.2011.08.030
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发表时间：
2011-10-30
影响因子：
3
通讯作者：
Sabbagh MN
中科院分区：
文献类型：
--
作者：
Gonzales A;Decourt B;Walker A;Condjella R;Nural H;Sabbagh MN
乙酰基蛋白酶BACE 1是β淀粉样蛋白合成的限速酶，β淀粉样蛋白在人脑中的积累是阿尔茨海默病（AD）的标志。BACE 1已被提议作为AD的替代标志物;然而，很少有BACE 1免疫测定的报道。在本研究中，我们通过Western blot和几个抗体对筛选了10个BACE 1抗体，建立了一种新的BACE 1夹心ELISA方法。我们确定了一对，表现出很少的背景和良好的再现性。尝试了几种稀释缓冲液和样品变性方法以在捕获前部分解折叠BACE 1。我们发现，在PBS中稀释，然后在50 °C下孵育10分钟，显著提高了测定的性能。最后，我们成功地测量了几个人脑和血小板裂解物以及血浆和AD CSF中的BACE 1水平。我们预计，这种方法将奠定基础，准确地测量BACE 1在人体组织中的水平，这可能有助于在不久的将来AD的分子诊断。
The aspartyl protease BACE1 is the rate limiting enzyme in the synthesis of amyloid beta, which accumulation in the human brain is a hallmark of Alzheimer’s disease (AD). BACE1 has been proposed as a surrogate marker of AD; however, very few BACE1 immunoassays have been reported. In the present study we have screened ten BACE1 antibodies by Western blot and several antibody pairs to develop a new BACE1 sandwich ELISA procedure. We identified one pair that showed little background and good reproducibility. Several dilution buffers and sample denaturation methods were tried to partially unfold BACE1 before capture. We found that dilution in PBS followed by 10 min incubation at 50 °C critically improves the performance of the assay. Finally, we successfully measured BACE1 levels in a few human brain and platelet lysates as well as in plasma and AD CSF. We anticipate that this assay will lay the ground to accurately measure BACE1 levels in human tissues, which could facilitate the molecular diagnosis of AD in the near future.
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DOI：
10.1523/jneurosci.3657-09.2009
发表时间：
2009-10-14
期刊：
The Journal of neuroscience : the official journal of the Society for Neuroscience
影响因子：
--
作者：
Vassar R;Kovacs DM;Yan R;Wong PC
通讯作者：
Wong PC
影响因子：
4.8
作者：
Kalvodova, L;Kahya, N;Simons, K
通讯作者：
Simons, K
影响因子：
4.8
作者：
von Arnim, CAF;Kinoshita, A;Hyman, BT
通讯作者：
Hyman, BT
影响因子：
9.3
作者：
Verheijen, Jan H.;Huisman, Linda G. M.;Hanemaaijer, Roeland
通讯作者：
Hanemaaijer, Roeland
影响因子：
3.5
作者：
Hussain, I;Powell, D;Christie, G
通讯作者：
Christie, G