蛋白质芯片技术介绍

1、The future is in your hands!Nowadays,the poteomics(蛋白质的)research is one of a general direction of the development of life science after genomics（基因组学的）research. Protein structure and function have the direct impact on the change of life activity.And the level of gene transcription(转录) can only refle

2、cts the change of gene expression products in a certain extent. However, truly functional protein processes after the transcription, translation regulation processing ,controling to form and so on steps. Thus the direct study of protein to the interpretation(翻译) of the real life phenomenonis of grea

3、t improtance. But research on means and methods(手段方法) of the protein doesnt have a big development, so the search for effective and fast protein analysis technology has become one of the most important step.Then comes the protein chip The protein chip（蛋白芯片） technology is quickly developed in recent

4、years in chemical biology as a high and new technology.It is the basic principle is make various proteins orderly fixed on all kinds of medium carrier（介质载体） as a test chip.And then, using the tag antibodies(抗体) that are specified by the fluorescent(荧光) substance match with the protein.And the corres

5、ponding(对应的)protein will matchwith the chip. Then,the fluorescent on the antibody will indicate the corresponding(对应的) number of protein and its expression. All the other antibodies that doesnt make complementary combination will be washed after using the fluorescent scanner(荧光扫描仪)or laser scanning

6、technology(激光扫描共聚技术).Through analysisthe fluorescence intensity(强度)of each point on the chip and interaction relationship between protein and protein, the purpose of achieving determining all sorts of function of gene expression will be done.There are three types of protein microarrays that are curr

7、ently used to study the biochemical activities of proteins.Analytical microarrays are also known as capture arrays. In this technique, a library of antibodies, aptamers or affibodies is arrayed on the support surface. These are used as capture molecules since each binds specifically to a particular

8、protein. The array is probed with a complex protein solution such as a cell lysate. Analysis of the resulting binding reactions using various detection systems can provide information about expression levels of particular proteins in the sample as well as measurements of binding affinities and speci

9、ficities. This type of microarray is especially useful in comparing protein expression in different solutions. For instance the response of the cells to a particular factor can be identified by comparing the lysates of cells treated with specific substances or grown under certain conditions with the

10、 lysates of control cells. Another application is in the identification and profiling of diseased tissues.Functional protein microarrays (also known as target protein arrays) are constructed by immobilising large numbers of purified proteins and are used to ideantify protein-protein, protein-DNA, pr

11、otein-RNA, protein-phospholipid, and protein-small molecule interactions, to assay enzymatic activity and to detect antibodies and demonstrate their specificity. They differ from analytical arrays in that functional protein arrays are composed of arrays containing full-length functional proteins or

12、protein domains. These protein chips are used to study the biochemical activities of the entire proteome in a single experiment.Reverse phase protein microarray (RPPA) involve complex samples, such as tissue lysates. Cells are isolated from various tissues of interest and are lysed. The lysate is ar

13、rayed onto the microarray and probed with antibodies against the target protein of interest. These antibodies are typically detected with chemiluminescent, fluorescent or colorimetric assays. Reference peptides are printed on the slides to allow for protein quantification of the sample lysates. RPAs

14、 allow for the determination of the presence of altered proteins or other agents that may be the result of disease. Specifically, post-translational modifications, which are typically altered as a result of disease can be detected using RPAs.The advantages of protein chip1.All of the coarse biologic

15、al samples(粗生物样品) can be directly analyzed (serum, urine, body fluids) 直接用粗生物样品（血清、尿、体液）进行分析2.It can rapidly detect multiple biomarkers at the same time 同时快速发现多个生物标记物3. A small amount of sample只需要小量样品（约2-10L）4. High flux ability of validation高通量的验证能力（一次性验证多个蛋白质分子） found low abundance proteins发现低丰度蛋白

16、质（即在总蛋白提取物中含量很少的蛋白）6. Determinate the hydrophobic protein测定疏水蛋白质(疏水蛋白质具有不同寻常的生化性质, 很难用常规的蛋白分析方法检测到其存在)7.In the same system,make cimc discovery and detection all together.Using monoclonal antibody chip to identify the unknown antigen/protein, to reduce the determination of protein sequence of work. 在

17、同一系统中集发现和检测为一体利用单克隆抗体芯片，鉴定未知抗原/蛋白质，以减少测定蛋白质序列的工作量。8.It can quantitatively use monoclonal antibody chip,.As a result of that the combination to chip antibody is quantitative, it can be used to measure quantity of antigen.可以定量利用单克隆抗体芯片，由于结合至芯片上的抗体是定量的，故可以测定抗原量.9. Using monoclonal antibody microarrays

18、can comple the function of insufficient cells,such as dissolve cells, determinating the intracellular antigens.Whats more,its sensitivity is much higher than that.利用单克隆抗体芯片,可互补流式细胞仪不足的功能，如将细胞溶解，可测定细胞内的抗原，而且灵敏度远高于流式细胞仪。There are five major areas where protein chips are being applied: diagnostics(诊断),

19、 proteomics, protein functional analysis, antibody characterization, and treatment development.Diagnostics involves the detection of antigens and antibodies in blood samples; the profiling of sera to discover new disease biomarkers; the monitoring of disease states and responses to therapy in person

20、alized medicine; the monitoring of environment and food.Proteomics pertains to protein expression profiling i.e. which proteins are expressed in the lysate of a particular cell.Protein functional analysis is the identification of protein-protein interactions (e.g. identification of members of a prot

21、ein complex), protein-phospholipid interactions, small molecule targets, enzymatic substrates (particularly the substrates of kinases) and receptor ligands.Antibody characterization is characterizing cross-reactivity, specificity and mapping epitopes.Treatment development involves the development of

22、 antigen-specific therapies for autoimmunity, cancer and allergies; the identification of small molecule targets that could potentially be used as new drugs.(1) Establishing a fast, cheap, high protein expression and purification method of flux, high flux and defining each antibody preparation of antibody affinity specificity; The first generation of protein detection chip will rely mainly on antibodies and other macromolecules, clearly, with these material preparation complex chips, scale production will exist a lot of practical prob