Construction of CTB fusion proteins for screening of monoclonal antibodies against Salmonella typhi OmpC peptide loops.
Mice were immunized with resin-bound peptides whose sequences have been proposed to be part of exposed loops in Salmonella typhi outer membrane protein OmpC. To screen hybridomas for monoclonal antibodies against those epitopes, we designed fusion proteins where the candidate peptide sequence was attached to the amino end of cholera toxin B-subunit (CTB). The constructed fusion proteins allowed the efficient selection of positive clones by GM1-ELISA. Selected antibodies recognized purified OmpC and whole Salmonella bacteria. This suggests a native structure of our genetically attached peptides in agreement with immunological properties reported for previous CTB recombinant fusion proteins. In a more general context, CTB hybrids could be used to screen for antibodies towards immunogenic epitopes in other systems. This might turn out to be particularly useful when producing antibodies against peptide sequences in microorganisms whose handling is difficult or that pose inherent health risks.
Paniagua-Solís
J
J
Unidad de Investigación Médica en Immunoquímica, Hospital de Especialidades, Instituto Mexicano del Seguro Social, Mexico City, Mexico.
Sánchez
J
J
Ortiz-Navarrete
V
V
González
C R
CR
Isibasi
A
A
D000595:Amino Acid Sequence; D000818:Animals; D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins / Q000032:analysis / Q000235:genetics / Q000276:immunology*; D001483:Base Sequence; D002772:Cholera Toxin / Q000235:genetics / Q000276:immunology*; D000939:Epitopes / Q000032:analysis; D006825:Hybridomas; D051379:Mice; D008809:Mice, Inbred C3H; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology; D012485:Salmonella typhi / Q000235:genetics / Q000276:immunology*
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins; D000939:Epitopes; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D002772:Cholera Toxin