title: Preferential labeling of alpha-amino N-terminal groups in peptides by biotin: application to the detection of specific anti-peptide antibodies by enzyme immunoassays.
abstract: Experimental conditions (pH 6.5, 24 h reaction, peptide:biotin ratio 1:5) were defined for preferential incorporation of the biotin molecule in the N-terminal alpha-amino group of peptides. This strategy could be helpful in numerous applications when an entire peptide chain must remain accessible for antibody or receptor binding. We illustrate this advantage in a solid-phase enzyme immunoassay designed to detect antibodies specific for bovine beta-lactoglobulin present in rabbit or human sera. This test involves synthetic peptides biotinylated in different positions and immobilized on a solid phase. The use of biotin/streptavidin interactions permitted more efficient detection of specific anti-peptide antibodies than solid phases prepared using conventional passive-adsorption techniques. The highest levels of antibody binding were measured when biotinylation occurred at the N-terminal extremity of immobilized peptides.
authors.affiliation: Laboratoire Associé INRA-CEA d'Immuno Allergie Alimentaire, SPI Bât 136, Gif sur Yvette, France.
authors.forename: I
authors.initials: I
authors.lastname: Sélo
authors.forename: L
authors.initials: L
authors.lastname: Négroni
authors.forename: C
authors.initials: C
authors.lastname: Créminon
authors.forename: J
authors.initials: J
authors.lastname: Grassi
authors.forename: J M
authors.initials: JM
authors.lastname: Wal
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001710:Biotin / Q000737:chemistry*; D002417:Cattle; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007124:Immunoenzyme Techniques; D007782:Lactoglobulins / Q000276:immunology; D008894:Milk Proteins / Q000276:immunology; D008969:Molecular Sequence Data; D010455:Peptides / Q000737:chemistry*; D011817:Rabbits
chemical_list: D007782:Lactoglobulins; D008894:Milk Proteins; D010455:Peptides; D001710:Biotin