Phage-displayed random peptide libraries have been widely used for the selection of ligands that are able to bind homogeneous ligates, such as monoclonal antibodies (MAbs) or other proteins. Developing the strategy of using sera, for the selection of phage-displayed libraries, has imparted several very important advantages; when the sera are of pathological interest, it could lead to the discovery of disease-related epitopes and these could have diagnostic and/or prognostic value and could even be exploited for identifying unknown etiologic agents. This chapter describes a general strategy in identifying disease-specific phagotopes that avails itself only of the clinically characterized sera from patients and normal individuals. This strategy involves a three-step protocol: affinity selection, with one disease-specific serum, immunoscreening of the selected phage mixture, with other patient's sera, and counterscreening, using many different sera not related to the specific disease. The phage clones that are not recognized by all or most of the “negative” sera and yet react with the antibodies contained in the patients' sera can, thus, be identified and characterized. This entire process is, therefore, workable without the requirement of or even information about the etiologic agent or its antigens.
Phage-displayed peptides as tools for the characterization of human sera
FELICI, Franco;
1996-01-01
Abstract
Phage-displayed random peptide libraries have been widely used for the selection of ligands that are able to bind homogeneous ligates, such as monoclonal antibodies (MAbs) or other proteins. Developing the strategy of using sera, for the selection of phage-displayed libraries, has imparted several very important advantages; when the sera are of pathological interest, it could lead to the discovery of disease-related epitopes and these could have diagnostic and/or prognostic value and could even be exploited for identifying unknown etiologic agents. This chapter describes a general strategy in identifying disease-specific phagotopes that avails itself only of the clinically characterized sera from patients and normal individuals. This strategy involves a three-step protocol: affinity selection, with one disease-specific serum, immunoscreening of the selected phage mixture, with other patient's sera, and counterscreening, using many different sera not related to the specific disease. The phage clones that are not recognized by all or most of the “negative” sera and yet react with the antibodies contained in the patients' sera can, thus, be identified and characterized. This entire process is, therefore, workable without the requirement of or even information about the etiologic agent or its antigens.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
