Proteins can be immobilized (active but not soluble) in a number of different ways. When a protein is covalently coupled to an insoluble support, two techniques may be employed. One technique is to activate the support material for reaction with groups on the protein. Second, one can use a coupling reagent to link protein to matrix. A third possibility would be to activate the protein molecule for coupling to the support material. This chapter discusses the groups on the protein that is available for covalent coupling to various matrices and some of the most commonly used procedures. The reactivity of a certain group in the protein depends on the location and environment in which the amino acid residues are placed. Because the proteins are large molecules, many of the site changes are partially “buried” and may be relatively inert, while some residues that are exposed to the solvent may be highly reactive. The principles discussed in the chapter concerning the reaction of the amino acid residues on protein molecules with an active group on some support material, are also applicable to the other methods involving covalent coupling of the protein.
ASJC Scopus subject areas
- Molecular Biology