基因,蛋白,免疫 课程之免疫学.doc

Monoclonal antibodies - agents for structural analysis References The molecular dissection of protein antigens by MHV Van Regenmortal in Structure of Antigens vol 1. CRC Press 1992. MAbs as aids in structural analysis Antibodies carry antigen specific combining sites or paratopes which recognize epitopes on the surface of protein molecules. Carbohydrate, lipid and nucleic acids can also carry epitopes. Monoclonal antibodies (MAbs) bind to a single epitope which, by appropriate techniques, can be identified within the amino acid sequence of the target protein. Mabs can therefore be used to probe the surface of a folded protein. As binding of MAbs may influence function, they may also be used to investigate structure/function relationships. 1. MAbs raised against a complex antigen may be used to differentiate and identify component parts of the antigen. 2. Epitope mapping of MAbs selected for functional activity can identify intra-molecular structures responsible for that function. 3. MAbs raised against a single protein or epitope may be used to locate or track that protein. 4.4. MAbs can also be used to monitor changes in protein structure as for instance in protein folding and refolding. Linear vs conformational epitopes Linear or continuous epitopes are short linear sequences of amino acids which are able bind antibody raised against the native protein from which they were derived. The amino acid sequence may be presented to the antibody as a peptide fragment or within the denatured, unfolded polypeptide chain of the whole protein. Such sequences are generally 5-8 amino acids in length. A conformation-dependent or discontinuous epitope is comprised of amino acids which are not contiguous in the polypeptide chain but are brought together on the surface of the folded protein. The full discontinuous epitope appears to involve approximately 20 amino acids. Continuous epitopes may be distinguished from discontinuous by their ability to bind to proteins lineariz