新浪登录
腾讯登录蛋白生物标志物的筛选
| 导读 | 肿瘤、代谢病和心血管疾病等，尤其是肿瘤，是一个有很多因素参与的病理过程，只对其中的一种或几种蛋白进行检测，无法对疾病进行整个系统的研究。在生物标志物发现初期······ |
肿瘤、代谢病和心血管疾病等，尤其是肿瘤，是一个有很多因素参与的病理过程，只对其中的一种或几种蛋白进行检测，无法对疾病进行整个系统的研究。在生物标志物发现初期，应该对多种可能与疾病相关的蛋白或分子进行全面筛选。传统的单因子检测方法已不能满足科学工作者日益增多的筛选需求，而高通量检测工具可同时对多种蛋白或分析物进行检测，实现了高通量筛选和定量的目的，广泛应用于疾病标志物的早期筛查。
为什么使用高通量多因子检测技术？· 单一样本量太少，或样本太珍贵，无法支持多个指标的检测时-节省样本
· 样本太多，想快速转化为具有临床意义的数据，不想没完没了的做实验时-省时省力
· 老板给的经费太少了，我该如何能用最少的钱得到最多的结果时-节约开支
· 指标太多了，哪一个跟我的实验表型最相关，需要筛选时-高通量筛选
我们提供多种高通量筛选工具：
1. 固相抗体芯片
2. Luminex®液相抗体芯片
3. Tocriscreen™小分子化合物库
1. 固相抗体芯片
Western Blot一直是蛋白半定量检测的常用技术，然而Western Blot需要进行制胶、电泳、转膜、抗体孵育、显色和成像，操作过程复杂且需要进行条件优化，得到一次较好的实验结果耗时良久。R&D Systems公司提供的Proteome Profiler™固相芯片，能达到和WB相同的实验结果，但却无需繁琐的实验步骤，一次即可实现最多达百种以上蛋白检测，成为蛋白半定量的最佳筛选工具。
特点
· 重要文献中引用次数近700次
· 可同时检测上百种分析物
· 比Western blot更简单，更灵敏
· 无需特殊实验设备
· 手工操作时间为3小时
检测原理
应用
1) 筛选乳腺癌磷酸化蛋白
查看更多内容请点击>>
文献
1.Hemant K. Bid, Ryan D. Roberts, et al. △Np63 Promotes Pediatric Neuroblastoma and Osteosarcoma by Regulating Tumor Angiogenesis. Cancer Res, 74(1) January 1, 20142.Zemskova, M., Song, J., Cen, B. et al. Regulation of prostate stromal fibroblasts by the PIM1 protein kinase. Cell Signal. 2015 Jan;27(1):135-46. [引用产品货号：ARY005, ARY001]
3.Cho YL, Hur SM, et al. Specific activation of insulin-like growth factor-1 receptor by ginsenoside Rg5 promotes angiogenesis and vasorelaxation. J Biol Chem. 2015 Jan 2;290(1):467-77. [引用产品货号：ARY003B]
4.Lalani, A., Moore, C. et al. Myeloid cell TRAF3 regulates immune responses and inhibits inflammation and tumor development in mice. J Immunol. 2015 Jan 1;194(1):334-48. [引用产品货号：ARY006]
5.Ariestanti, D., Ando, H., et al. Targeted Disruption of Ig-Hepta/Gpr116 Causes Emphysema-like Symptoms That Are Associated with Alveolar Macrophage Activation.J Biol Chem. 2015 Apr 24;290(17):11032-40. [引用产品货号：ARY006]
6.Wang, J., Mikse, O., et al. Ligand-associated ERBB2/3 activation confers acquired resistance to FGFR inhibition in FGFR3-dependent cancer cells. Oncogene. 2015 Apr 23;34(17):2167-77. [引用产品货号：ARY001B]
7.Man, J., Shoemake, J. et al. Hyperthermia Sensitizes Glioma Stem-like Cells to Radiation by Inhibiting AKT Signaling. Cancer Res. 2015 Apr 15;75(8):1760-9. [引用产品货号：ARY003B]
8.Annemann, M., Wang, Z. et al. IkappaBNS regulates murine Th17 differentiation during gut inflammation and infection. J Immunol. 2015 Mar 15;194(6):2888-98.[引用产品货号：ARY006]
9.Montero, J., Esparis-Ogando, A., et al. Active kinase profiling, genetic and pharmacological data define mTOR as an important common target in triple-negative breast cancer. Oncogene. 2014 Jan 9;33(2):148-56. [引用产品货号：ARY001B]
10.Stommel, JM., Kimmelman, AC., et al. Coactivation of receptor tyrosine kinases affects the response of tumor cells to targeted therapies. Science. 2007 Oct 12;318 (5848):287-90. [引用产品货号：ARY001]
11.Engelman, JA., Zejnullahu, K., et al. MET amplification leads to gefitinib resistance in lung cancer by activating ERBB3 signaling. Science. 2007 May 18;316 (5827):1039-43. [引用产品货号：ARY001]
2. Luminex液相抗体芯片
ELISA实验能够很好的对蛋白进行精确定量且操作简单，然而需要同时检测多个样本的多个因子时，ELISA需要多次重复相同的步骤，不能很快的得到实验结果。Lumimex液相芯片，以ELISA三明治夹心原理，利用微球和基于流式细胞仪原理的Lumimex仪器相结合的技术，采用ELISA板的形式实现多样本验证，在单孔中同时检测多个蛋白指标，进行精确定量。Luminex液相抗体芯片技术采用不同颜色编码的微球，包被相应的特异性抗体，在一个孔可同时检测多达100种分析物，利用96孔板作为反应板，实现高通量的蛋白精确定量。R&D Systems公司有15年Luminex试剂盒的研发历史，具有丰富的免疫检测经验，采用与ELISA试剂盒质量一致的抗体对和缓冲液体系优化，在“金标准ELISA”的基础上，提供最高标准和最优性能的Luminex多重检测试剂，对多个生物标志物进行定量和筛选。
特点
灵活：根据客户需求，可同时检测多达 100 种指标物
性价比高：相比于单因子检测，成本更低
选择性多：可检测 的指标种类数不断增加，部分因子是 R&D
Systems 公司独有
实验操作简单：整个实验流程在 3～3.5 小时内完成
同时检测多个指标所需的样本量极少（<50 μL）
实验原理
Fig. 1 Luminex 液相抗体芯片实验原理
将能识别不同目标分析物的微珠混合后与待检样本孵育，微珠捕获到分析物后，经由多个生物素标记的特异性检测抗体混合物识别，再与链霉亲和素-PE 荧光素结合后进行检测。
应用
文献：
1.Anaïs Levescot, Stéphane Flamant, et al. BCR-ABL–Induced Deregulation of the IL-33/ST2 Pathway in CD34(+) Progenitors from Chronic Myeloid Leukemia Patients.Cancer research. 2014 May 15; 74: 2669.
2.Myers, N., Chantratita, N., et al. The role of NOD2 in murine and human melioidosis. J Immunol. 2014 Jan 1; 192(1): 300-7.
3.Dubsky, M., Jirkovska, A., et al. Role of serum levels of angiogenic cytokines in assessment of angiogenesis after stem cell therapy of diabetic patients with critical limb ischemia. Cell Transplant. 2014; 23(12): 1517-23.
4.Garg, , Abhishek., Dudek, Aleksand., et al. ROS-induced autophagy in cancer cells assists in evasion from determinants of immunogenic cell death. Autophagy. 2013 Sep; 9(9): 1292-307.
5.David M., Charytan, Robert Padera, et al. Increased concentration of circulating angiogenesis and nitric oxide inhibitors induces endothelial to mesenchymal transition and myocardial fibrosis in patients with chronic kidney disease. J Immunol. 2014 Sep; 176(1): 99-109.
6.Richard Fuerstenberg, Shaya Anderson, et al. Multiplex analysis of secreted biomarkers from stimulated natural killer cells (INC6P.337). J Immunol. 2014 Sep; 176(1):99-109. 2014 May 1; 192(1): Supplement 121.4.
7.Yurkovetsky Z., Skates S., et al. Development of a multimarker assay for early detection of ovarian cancer. J. Clin. Oncol. 2010; 28(13): 2159-66.
8.Nolen B., Breen E., et al. Circulating mediators of inflammation and immune activation in AIDS-related non-hodgkin lymphoma. PLoS ONE. 2014; 9(6): e99144.
9.Grote VA., Kaaks R., et al. Inflammation marker and risk of pancreatic cancer: a nested case-control study within the EPIC cohort. Br. J. Cancer. 2012; 106(11): 1866-74.
10.Chantratita N, Tandhavanant S., et al. Survey of innate immune responses to Burkholderia pseudomallei in human blood identifies a central role for lipopolysaccharide. PLoS ONE. 2013; 8(11): e81617.
11.Madsen C, Steffensen K., et al. Serial measurements of serum PDGF-AA, PDGF-BB, FGF2, and VEGF in multiresistant ovarian cancer patients treated with bevacizumab.JOvarian Res. 2012; 5(1): 23.
12.Chantrathammachart, P. Mackman, N., et al. Tissue factor promotes activation of coagulation and inflammation in a mouse model of sickle cell disease. Blood. 2012 Jul 19;120(3): 636-46.
13.Vuillermot, S. Joodmardi, E., et al. Prenatal immune activation interacts with genetic Nurr1 deficiency in the development of attentional impairments. J Neurosci. 2012 Jan 11;32(2): 436-51
2.Myers, N., Chantratita, N., et al. The role of NOD2 in murine and human melioidosis. J Immunol. 2014 Jan 1; 192(1): 300-7.
3.Dubsky, M., Jirkovska, A., et al. Role of serum levels of angiogenic cytokines in assessment of angiogenesis after stem cell therapy of diabetic patients with critical limb ischemia. Cell Transplant. 2014; 23(12): 1517-23.
4.Garg, , Abhishek., Dudek, Aleksand., et al. ROS-induced autophagy in cancer cells assists in evasion from determinants of immunogenic cell death. Autophagy. 2013 Sep; 9(9): 1292-307.
5.David M., Charytan, Robert Padera, et al. Increased concentration of circulating angiogenesis and nitric oxide inhibitors induces endothelial to mesenchymal transition and myocardial fibrosis in patients with chronic kidney disease. J Immunol. 2014 Sep; 176(1): 99-109.
6.Richard Fuerstenberg, Shaya Anderson, et al. Multiplex analysis of secreted biomarkers from stimulated natural killer cells (INC6P.337). J Immunol. 2014 Sep; 176(1):99-109. 2014 May 1; 192(1): Supplement 121.4.
7.Yurkovetsky Z., Skates S., et al. Development of a multimarker assay for early detection of ovarian cancer. J. Clin. Oncol. 2010; 28(13): 2159-66.
8.Nolen B., Breen E., et al. Circulating mediators of inflammation and immune activation in AIDS-related non-hodgkin lymphoma. PLoS ONE. 2014; 9(6): e99144.
9.Grote VA., Kaaks R., et al. Inflammation marker and risk of pancreatic cancer: a nested case-control study within the EPIC cohort. Br. J. Cancer. 2012; 106(11): 1866-74.
10.Chantratita N, Tandhavanant S., et al. Survey of innate immune responses to Burkholderia pseudomallei in human blood identifies a central role for lipopolysaccharide. PLoS ONE. 2013; 8(11): e81617.
11.Madsen C, Steffensen K., et al. Serial measurements of serum PDGF-AA, PDGF-BB, FGF2, and VEGF in multiresistant ovarian cancer patients treated with bevacizumab.JOvarian Res. 2012; 5(1): 23.
12.Chantrathammachart, P. Mackman, N., et al. Tissue factor promotes activation of coagulation and inflammation in a mouse model of sickle cell disease. Blood. 2012 Jul 19;120(3): 636-46.
13.Vuillermot, S. Joodmardi, E., et al. Prenatal immune activation interacts with genetic Nurr1 deficiency in the development of attentional impairments. J Neurosci. 2012 Jan 11;32(2): 436-51
3. Tocriscreen™小分子化合物库
Tocriscreen collections是独特的小分子库，适用于基于化学遗传学、化学生物学的筛选、受体脱孤化、靶点验证或药物再分析。由Tocris目录中精选的生物活性化合物所组成，Tocriscreen collections全面覆盖了生物化学和细胞筛选检测中最重要的靶点。它们可用于高通量和高内涵筛选，为现代药物开发提供了一个不可或缺的起点。这些库中的小分子预溶解在DMSO中，包含了针对GPCR、激酶、离子通道、核受体、转运蛋白和干细胞生物学的化合物。许多化合物无法从别的途径获得。
特点
● 所有化合物都具有生物活性
● 可提供完整的化学和生物学数据
● 许多化合物都是 Tocris 独有的
● 非同一般的纯度
● 保证化合物的再次供应
（转化医学网360zhyx.com）
还没有人评论，赶快抢个沙发