Affinity chromatography

Examples of Affinity chromatography in the following topics:

• Mapping Protein-Protein Interactions

  • Affinity purification of protein complexes coupled to mass spectrometry is carried out as follows: a specific protein (the bait) is manipulated to express an affinity tag.
  • The tag serves as a tool to purify the bait protein and associated proteins by affinity chromatography.

• Purifying Proteins by Affinity Tag

  • Affinity tags are appended to proteins so that they can be purified from their crude biological source using an affinity technique.
  • Chromatography tags are used to alter chromatographic properties of the protein to afford different resolution across a particular separation technique.

• Metabolomics

  • The term "metabolic profile" was introduced by Horning, et al. in 1971, after they demonstrated that gas chromatography-mass spectrometry (GC-MS; ) could be used to measure compounds present in human urine and tissue extracts.
  • GC-MS is a method that combines the features of gas-liquid chromatography and mass spectrometry to identify different substances within a test sample.
  • Separation methods: Gas chromatography, especially when interfaced with mass spectrometry (GC-MS), is one of the most widely used and powerful methods.
  • Gas chromatography–mass spectrometry (GC-MS) is a method that combines the features of gas-liquid chromatography and mass spectrometry to identify different substances within a test sample.

• Northern Blots

  • Eukaryotic mRNA can then be isolated through the use of oligo (dT) cellulose chromatography to isolate only those RNAs with a poly(A) tail.

• Crystallographic Analysis

  • .), purification of recombinant proteins (such as chromatography of affinity and gel filtration), enzymatic tests and inhibition measurement (spectrophotometry), crystallization, x-rays crystallography and structural analysis, interactions determination (microcalorimetry, fluorescence, BIAcore), conformational analyses (circular dichroism, ultracentrifugation, light scattering), modifications analysis (mass spectrometry), bioinformatics, and molecular modelisation.

• Antibody Genes and Diversity

  • Affinity maturation occurs after V(D)J recombination, and is dependent on help from helper T cells.

• Quorum Sensing

  • Recently, a well-studied quorum quenching bacterium has been isolated and its AHL degradation kinetic has been studied by using rapid resolution liquid chromatography (RRLC).

• Proteomics

  • New methods include protein microarrays, immunoaffinity, and chromatography followed by mass spectrometry , dual polarisation interferometry, Microscale Thermophoresis, and experimental methods such as phage display and computational methods.

• Obtaining DNA

  • The DNA is then purified using simple methods to remove contaminating proteins (extraction with phenol), RNA (ribonuclease) and smaller molecules (precipitation and/or chromatography).