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Healthier biomarkers are without delay necessary to progress finding the disease. Biomarker is used to identify and measure the stage of disease from the specific biological samples. Molecular biomarkers are formed in many types for different strategies implement for new discovery. New biomarker discovery shown well-built prospective to identify cancer, diabetics, genetic disorder and metabolic ailment and also identify the toxicity studies with drug . Protein sphere of influence is probable for the mainly all over the place pretentious in disease and new biomarker focus in development of new protein biomarker.
Henry Bence- Sones start first cancer biomarker protein study in 1847 . Main activity of biomarker is used for clinical purpose to screen the slight movement of disease or diagnosis and identify the molecular objective therapy. It is very useful to identify the specific disease in early stage diagnosis. Most of the bio pharmaceutical industries focus to projection of novel biomarker with the help of molecular method . The identification of novel biomarker travel lengthy and hard path to find applicant finding to clinical assay, and the short of coherent and wide-ranging pipeline for development of new biomarker. Biomarker discovery and validation based on understand the specific or target protein function and coherent study. Processes of biomarker discover and validation by different pipeline like quantification of sample, discovery of novel biomarker, verification of the sample and finally clinical validation by the cohort study.
The Human Protein Atlas is one the best opportunity used to analysis of antibody-based proteomics in perfect approach for the identification of new biomarker. It is open database contain collection of high quality image showing the allocation of different biological sample like human tissue, different type of cancer and cell line. The Human protein atlas database large number of appliance for specific antibody validation with the help of western blot, immunohistochemisty, protein array, immunofluorescent analysed by confocal microscopy. The human protein atlas latest version 8 released on 2011 may 16, it contain 11260 genes with protein expression profiles based on 14506 antibodies. The first human protein atlas database version 1 released on 2005 contain 413568 images and only 718 antibodies present.
The antibody validation and the high throughput approach based in the human protein atlas development used look for probable clinical biomarker from the database. In the human protein atlas contain advance search tool used to identifying the specific biomarker from the large database based on cell, tissue, cancer, malignancy specific biomarker, prognostic and diagnostic biomarker for specific disease identify by using advance search tool. We are focused in colorectal cancer based by group . Colorectal cancer is a bowel cancer, categorized by neoplasia in rectum and colon. It is most common cancer in the world and it kills annually more than 600 thousand people .
Cancer biomarkers can be used to selection test for colorectal cancer diagnosis. The factor gives explanation for the cause of molecular and protein expression level of colorectal cancer. At present biomarker is urgent need to resolve disease with the help of diagnosis biomarker. Colorectal biomarker divided in three types based on their sample tissue, stool and serum based method , but most of the analysis method used with the serum and tissue based sample. Normal to identify the new biomarker discovery is based on different channels, it divided into phase, sample, process, number of analyses and number of sample. In phase contain discovery, qualification, verification and antibody validation. Samples isolated from the serum sample is gold standard is reduced the biological variation from the different population sample analysis by the process of LC-MS/MS along with the immunoassay to identify the validation of the novel biomarker by the cohort study.
Figure 1: The method flow for the improvement of discovering the novel biomarker
Prognosis biomarker is differential expression within a cancer type and diagnosis biomarker is differing from the prognosis biomarker, it used for specific single cancer type . The human protein atlas database contains more detail related to cancer biomarker, advance tool used to filter from database by field, term and tissue type and expression level of the specific disease. In our search criteria we searched in the human protein atlas used to different search for colorectal cancer from the other data. Totally we identify 30 different genes from different filter level in that HPA 001042 and CAB011435 used for diagnostic biomarker. CAB 009454 and HPA 034661 identify used best for prognostic colorectal cancer. HPA 001042 indicate SATB2 colorectal expression level is 73% and CAB 011435 expressed in FASLG expression level is 82% gene used for diagnostic biomarker. Prognosis biomarker are CAB 009454 and HPA 034661 expressed level higher in moderate level 36 % compare to stronger level 26% expressed. Finally we selected only two biomarkers for each from prognosis and diagnosis biomarker
Result and Discussion:
Diagnostic and Prognostic biomarker:
Figure 2:CAB011435 for colorectal cancer diagnostic biomarker
Figure 3: Immunohistochemistry for differential staining for strong and negative staining
Figure 4: HPA 034661 is the prognostic biomarker
Figure 5: Immunohistochemistry differential staining indicate strong, medium and negative staining
Diagnostic biomarker used for specific single cancer type, here CAB011317 antibody expressed in FASLG gene used for colorectal cancer. FASLG gene is Fas ligand and it one of the tumour necrosis factor super family. It expressed 82% and 18 % weak in CAB011317 antibody and it validated by Immunohistochemistry staining of human tonsil sample illustrate different cytoplasmic and membranous positive result in a division of lymphoid cells and western blot result is perfect band is present in the antibody dilution of 1:5000. In the prognostic biomarker supported to HPA034661 for the differential expression within a cancer type, the gene encodes ARHGEF33 molecular function is Rho guanine nucleotide exchange factor. In the straining summary for immunofluorescent indicate negative result there is not even single strain observed in three cell lines, but in the Immunohistochemistry shows strong positive in cytoplasmic was experiential in Purkinje cells and negative result observed in normal tissues. Western blot is not shows specific band it expressed in weak band. Protein array expressed very perfectly with single peak corresponding to interface level.
Cohort analysis is used for size of control and case, complexity focus to age, sex, gender and ethical. It is very used to unbiased analyse discoveries of novel biomarker and validation is the final verification for the biomarker. So far only nine protein biomarker are approvable by FDA in 2011. So identification of new biomarker is in urgent need for the colorectal cancer.