Characterization of cerebrospinal fluid antibody specificities in neurocysticercosis using phage display peptide library.
To identify epitopes for antibodies present in cerebrospinal fluid (CSF) samples from two patients with confirmed neurocysticercosis, we used a phage peptide library that displayed random dodecapeptides as a fusion to the minor coat protein (pIII) of phage M13. To increase the specificity of selection, plates coated with anti-human Fc antibody were used in five rounds of biopanning. The DNA inserts of 30 selected clones were determined and the deduced amino acid sequences were analyzed. Sequence similarities between affinity-selected clones and three known T. solium and T. crassiceps proteins were encountered. Two phage clones have been shown to react in enzyme-linked immunosorbent assay with CSF samples from other patients with neurocysticercosis, and no reaction was observed with CSF samples from patients with other neurological disorders used as negative controls. Our results suggest that affinity selection of peptides from phage libraries using CSF samples from patients is an attractive tool for the development of novel reagents for a simple and sensitive immunodiagnostic test and for understanding the molecular mechanisms participating in the pathogenesis of neurocysticercosis.
Manoutcharian
K
K
Instituto de Investigaciones Biomedicas, Universidad Nacional Autonoma de Mexico, Mexico.
Sotelo
J
J
Garcia
E
E
Cano
A
A
Gevorkian
G
G
D000595:Amino Acid Sequence; D000818:Animals; D000909:Antibodies, Helminth / Q000134:cerebrospinal fluid*; D000918:Antibody Specificity; D000947:Antigens, Helminth / Q000737:chemistry / Q000235:genetics; D016022:Case-Control Studies; D003552:Cysticercus / Q000235:genetics / Q000276:immunology*; D000939:Epitopes / Q000737:chemistry / Q000235:genetics; D006801:Humans; D007159:Immunologic Tests; D020019:Neurocysticercosis / Q000175:diagnosis / Q000209:etiology / Q000276:immunology*; D019151:Peptide Library; D017386:Sequence Homology, Amino Acid
D000909:Antibodies, Helminth; D000947:Antigens, Helminth; D000939:Epitopes; D019151:Peptide Library