蛋白质芯片技术介绍
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——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 reflects 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 great improtance. But research on means and methods(手段方法) of the protein doesn't have a big development, so the search for effective and fast protein analysis technology has become one of the most important step.
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
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 arrayed 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 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.
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, protein-RNA, protein-phospholipid, and proteinsmall 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 protein domains. These protein chips are used to study the biochemical activities of the entire proteome in a single experiment.
在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.
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.
match with the protein.And the corresponding(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 doesn't make complementary combination will be washed after using the fluorescent scanner(荧光扫描仪)or laser scanning 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.
The advantages of protein chip
1.All of the coarse biological 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2-10μL） 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） ⒌ found low abundance proteins 发现低丰度蛋白质（即在总蛋白提取物中含量很少的蛋白）
Then comes the protein chip
The protein chip（蛋白芯片） technology is
quickly developed
in recent years in chemical
biology as a high and new technology.It is the basicΒιβλιοθήκη Baidu
There are three types of protein microarrays that are currently 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 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 specificities. 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 lysates of control cells. Another application is in the identification and profiling of diseased tissues.
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 reflects 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 great improtance. But research on means and methods(手段方法) of the protein doesn't have a big development, so the search for effective and fast protein analysis technology has become one of the most important step.
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
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 arrayed 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 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.
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, protein-RNA, protein-phospholipid, and proteinsmall 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 protein domains. These protein chips are used to study the biochemical activities of the entire proteome in a single experiment.
在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.
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.
match with the protein.And the corresponding(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 doesn't make complementary combination will be washed after using the fluorescent scanner(荧光扫描仪)or laser scanning 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.
The advantages of protein chip
1.All of the coarse biological 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2-10μL） 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） ⒌ found low abundance proteins 发现低丰度蛋白质（即在总蛋白提取物中含量很少的蛋白）
Then comes the protein chip
The protein chip（蛋白芯片） technology is
quickly developed
in recent years in chemical
biology as a high and new technology.It is the basicΒιβλιοθήκη Baidu
There are three types of protein microarrays that are currently 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 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 specificities. 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 lysates of control cells. Another application is in the identification and profiling of diseased tissues.