Generation of polyclonal antibodies against a chemically synthesized N-terminal fragment of the bacteriocin pediocin PA-1.
Six mice were immunized intraperitoneally (i.p.) with a chemically synthesized 9-mer fragment (PH1) designed from the N-terminal part of the bacteriocin pediocin PA-1 and conjugated to keyhole limpet haemocyanin (KLH). After three doses of the immunogen had been administered, serum-specific antibodies were detected by a competitive direct ELISA. Myeloma cells were injected i.p. into mice in order to obtain ascites polyclonal antibodies. Although four mice developed ascites, only mouse 2 had detectable specific antibodies in the ascites fluid. The serum and ascites antibodies were specific for PH1 but they did not recognize the whole pediocin PA-1 molecule. This is the first attempt to generate antibodies against bacteriocins with a chemically synthesized oligopeptide as immunogen. This approach still remains attractive for detection, quantification, mode of action studies and purification of bacteriocins, especially those for which the purification process is difficult or inefficient at present.
Martínez
M I
MI
Departamento de Nutrición y Bromatología III, Facultad de Veterinaria, Universidad Complutense de Madrid, Spain.
Rodríguez
J M
JM
Suárez
A
A
Martínez
J M
JM
Azcona
J I
JI
Hernández
P E
PE
D000595:Amino Acid Sequence; D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis*; D000918:Antibody Specificity; D000942:Antigens, Bacterial / Q000008:administration & dosage; D001430:Bacteriocins / Q000138:chemical synthesis / Q000235:genetics / Q000276:immunology*; D001667:Binding, Competitive; D016384:Consensus Sequence; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D007114:Immunization; D007274:Injections, Intraperitoneal; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D000070017:Pediocins; D010376:Pediococcus / Q000235:genetics / Q000276:immunology; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000235:genetics / Q000276:immunology*
D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001430:Bacteriocins; D000070017:Pediocins; D010446:Peptide Fragments; C074288:pediocin PA-1