Targeting procalcitonin with novel murine monoclonal antibodies.
The aim of this study was to produce monoclonal and polyclonal antibodies against procalcitonin (PCT), a valid post-mortem marker of sepsis marker. Monoclonal antibodies (MAbs) were made from hyperimmune Balb/c mice by injecting 50 microg of purified antigen. These mice were immunized four times and given a final boost, and their spleen cells were collected and fused with SP2/0 myeloma cells under the presence of PEG 1450. Hybridomas were screened by indirect enzyme-linked immunosorbent assay (ELISA) using purified protein. Lastly, five MAbs were obtained and designated successfully and were characterized by ELISA and Western immunoblotting. By Western immunoblotting, MAbs were found reactive with the patient's serum specifically. The results showed that the monoclonal antibodies could be used for various pathophysiological analyses in further investigations of procalcitonin and in preparing diagnostic kits.
Dong
Huimin
H
Institute of Antibody Engineering, School of Biotechnology, Southern Medical University, Guangzhou, China.
Shu
Wen
W
Liu
Tiancai
T
Wang
Suihai
S
Lin
Guanfeng
G
Li
Ming
M
Wu
Yingsong
Y
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology* / Q000302:isolation & purification; D000918:Antibody Specificity / Q000276:immunology*; D015153:Blotting, Western; D002116:Calcitonin / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D015740:Calcitonin Gene-Related Peptide; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D006825:Hybridomas; D051379:Mice; D008807:Mice, Inbred BALB C; D011498:Protein Precursors / Q000235:genetics / Q000276:immunology* / Q000302:isolation & purification; D011933:Reagent Kits, Diagnostic
D000911:Antibodies, Monoclonal; C000606589:CALCA protein, human; C000606590:Calca protein, mouse; D011498:Protein Precursors; D011933:Reagent Kits, Diagnostic; D002116:Calcitonin; D015740:Calcitonin Gene-Related Peptide