Finals notes - Protein Diagnostics/Detection o ELISA Enzyme Linked Immuno Sorbent Assay used to detect presence of antibody or antigen in a sample

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Protein Diagnostics/Detection o ELISA – Enzyme Linked Immuno Sorbent Assay used to detect presence of antibody or antigen in a sample Uses 2 antibodies; one binds to antigen and other to the antigen-Ab complexes and coupled to an enzyme First, fix known antigen to surface of plate; coat with serum Abs and wash; next add enzyme coupled second Abs; wash; apply substrate to elicit enzyme fluorescent signal; view with spectrophotometer o ImmunoPCR Immobilize an antigen to surface of plate; attach antibody- oligonucleotide complexes to antigen; this can now be detected by PCR due to the oligos by conductivity of fluorescent things DNA Diagnostics – in book o Hybridization with labeled DNA probe (label with biotin) Biotin recognized by streptovidin which is in turn linked to an enzyme and the DNA o Molecular beacons – when two dyes are close to each other they cancel each other’s signal, but if they hybridize you can see them Mutation Detection – requires little DNA o Sequence the DNA (isolate amplify sequence) Develop assays with high sensitivity to detect known sites of
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This note was uploaded on 12/13/2011 for the course BME 339 taught by Professor Georgiou during the Spring '11 term at University of Texas at Austin.

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Finals notes - Protein Diagnostics/Detection o ELISA Enzyme Linked Immuno Sorbent Assay used to detect presence of antibody or antigen in a sample

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