PROTEOMICS- PROTEIN MICROARRAY.pptx

Protein Microarray

1. SUBMITTED BY Ms. R. VAISHNAVI II M.Sc., Biotechnology PRIST UNIVERSITY Thanjavur SUBMITTED TO Dr. S. RAMADEVI Assistant Professor PROTEIN MICROARRAY

2. PROTEIN MICROARRY SYNOPSIS Introduction Types of array DNA micro array Protein microarray Protein microarray The probes on chip Sources of probes on chip Types of protein microarray Analytical protein microarray Function protein microarray A pplication of protein microarray

3. INTRODUCTION: Arrays are solid supports upon which a collection of gene- specific probes have been placed at defined locations, either by The term “micro” refers to the small size of the solid support. There are two major types of array DNA microarray Protein microarray Spotting synthesis Direct synthesis

4. PROTEIN MICROARRAY (A direct method for proteomics) A protein microarray (or protein chip) is a high- throughput method to track the interaction and activities of proteins, to determine their function, and determining function on a large scale. Its main advantage lies in the fact that large numbers of proteins can be tracked in parallel. THE PROBES ON CHIP A variety of materials can be immobilize on the protein chip based on the specific requirements. These include: Antibodies Antigens Aptamers Affibodies

5. THE SOURCES OF PROBES ON CHIP 1. Cell lysates: By using different chromatographic techniques like affinity chromatography, ion exchange chromatography etc. 2. Recombinant DNA Technology 3. Synthetic peptides: Solid- phase peptide synthesis.

6. TYPES OF PROTEIN MICROARRAY: 1. Analytical protein microarrays 2. Functional protein microarrays ANALYTICAL PROTEIN MICROARRAY Analytical microarrays are also known as capture arrays . 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 . Another applications is in the identification and profiling of diseased tissues.

7. FUNCTIONAL PROTEIN MICROARRAY Functional protein microarrays (also known as target protein arrays) are constructed by immobilizing large numbers of purified proteins and are used to identify protein- protein, protein- DNA, protein- RNA , and protein- phospholipid to assay enzymatic activity and to detect antibodies and demonstrate their specificity. These protein chips are used to study the biochemical activities of the entire proteome in a single experiment.

8. APPLICATIONS Diagnostics Proteomics Protein functional analysis Antibody characterization Treatment development DIAGNOSTICS Detection in antigen antibodies in blood To discover new disease biomarkers; monitoring the diseased state and responses to therapy. PROTEOMICS Protein expression profiling i.e., which proteins are expressed in a particular cell.

9. PROTEIN FUNCTIONAL ANALYSIS Protein- protein interactions Protein- phospholipid interactions Protein- DNA interactions Protein- RNA interactions Enzymatic substrates ANTIBODY CHARACTERIZATION characterization of cross reactivity, specificity and mapping epitopes. TREATMENT DEVELOPMENT Development of antigen- specificity therapies for autoimmunity, cancer and allergies. Identification of small molecules that could potentially used be as new drugs.