Immunoaffinity purifications

Immobilized antibodies may be used as affinity adsorbents for the antigens that stimulated their production (Figure 6.15). Antibodies, like many other biomolecules, may be immobilized on a suitable support matrix by a variety of chemical coupling procedures.

Immunoaffinity chromatography is amongst the most highly specific of all forms of biospe-cific chromatography. The high affinity with which an antibody normally binds its ligand can often make subsequent ligand desorption from the column difficult. Desorption can require conditions that result in partial denaturation of the bound protein. This is often achieved by alteration of buffer pH or by employing chemical disrupting agents, such as urea or guanidine. One of the most popular elution methods employed involves irrigation with a glycine-HCl buffer at pH 2.2-2.8. In some cases, elution is more readily attainable at alkaline pH values. Specific examples have been documented in which protein elution was performed under relatively mild conditions, such as a change of buffer system or an increase in ionic strength; however, such examples are exceptional. The inclusion of an immunoaffinity step in the purification of recombinant blood factor VIII used to treat haemophilia (Chapter 12) is one example of the industrial usage of this technique

Diabetes 2

Diabetes 2

Diabetes is a disease that affects the way your body uses food. Normally, your body converts sugars, starches and other foods into a form of sugar called glucose. Your body uses glucose for fuel. The cells receive the glucose through the bloodstream. They then use insulin a hormone made by the pancreas to absorb the glucose, convert it into energy, and either use it or store it for later use. Learn more...

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