Use of the phage display technique for detection of epitopes recognized by polyclonal rabbit gliadin antibodies.
A random phage heptapeptide library was screened with rabbit antibodies against wheat flour proteins comprising gliadins and a small amount of low molecular weight glutenins (gli/glu). Gli/glu antibodies isolated from the sera selected different consensus sequences (CS). All CS contained tri- to pentapeptide stretches homologous to gli/glu sequences (proposed epitopes). In alpha- and gamma-type gliadins, these sequences are clustered in the N-terminal region recently suspected to be toxic for humans with celiac disease. Peptides with CS were synthesized and checked for reactivity. Only immune and no control rabbit sera reacted with synthetic peptides. One of eight human sera containing gliadin antibodies was reactive as well (4/8 peptides) but control sera were negative. Thus the phage display technique is useful for epitope screening of polyclonal antibodies even in the case of a group of homologous but diverse antigens.
Osman
A A
AA
Institute of Clinical Chemistry and Pathobiochemistry, University of Leipzig, Germany.
Uhlig
H
H
Thamm
B
B
Schneider-Mergener
J
J
Mothes
T
T
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000276:immunology*; D001435:Bacteriophages; D016384:Consensus Sequence; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis* / Q000737:chemistry; D005903:Gliadin / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D000882:Haplorhini; D006801:Humans; D015151:Immunoblotting; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology; D019151:Peptide Library; D011817:Rabbits
D000906:Antibodies; D000939:Epitopes; D010446:Peptide Fragments; D019151:Peptide Library; D005903:Gliadin