TY - JOUR
T1 - Monoclonal antibody-based ELISA for detection of Clostridium perfringens alpha-toxin
AU - Liu, Zonglin L.
AU - Blaschek, Hans-Peter M
PY - 1996/1/1
Y1 - 1996/1/1
N2 - A monoclonal antibody-based ELISA was developed to detect alpha-toxin present in Clostridium perfringens bacterial cell lysates and cell- free culture supernatants. Monoclonal antibodies against C. perfringens alpha- toxin were produced in hybridoma tissue culture supernatants and in BALB/c mice ascites fluid. The monoclonal antibodies obtained from hybridoma culture supernatant and ascites fluid showed identical antigen specificity, but the latter showed a higher titer, with a 50% endpoint at 1/4,000. The monoclonal antibodies were specific for phospholipase C produced by C. perfringens, but not by Bacillus cereus. The lower limit of phospholipase C detection was 16 ng/ml. The dose-dependent relationship between absorption at 490 nm and concentration of phospholipase C diluted in HEPES (N- 2- hydroxyethylpiperazine-N'-2-ethanesulfonic acid) or Trypticase glucose yeast broth fit a four-parameter and a quadratic model, respectively. The monoclonal antibody-based ELISA developed is a rapid, sensitive and specific detection method and can be used for quantitative characterization of C. perfringens alpha-toxin.
AB - A monoclonal antibody-based ELISA was developed to detect alpha-toxin present in Clostridium perfringens bacterial cell lysates and cell- free culture supernatants. Monoclonal antibodies against C. perfringens alpha- toxin were produced in hybridoma tissue culture supernatants and in BALB/c mice ascites fluid. The monoclonal antibodies obtained from hybridoma culture supernatant and ascites fluid showed identical antigen specificity, but the latter showed a higher titer, with a 50% endpoint at 1/4,000. The monoclonal antibodies were specific for phospholipase C produced by C. perfringens, but not by Bacillus cereus. The lower limit of phospholipase C detection was 16 ng/ml. The dose-dependent relationship between absorption at 490 nm and concentration of phospholipase C diluted in HEPES (N- 2- hydroxyethylpiperazine-N'-2-ethanesulfonic acid) or Trypticase glucose yeast broth fit a four-parameter and a quadratic model, respectively. The monoclonal antibody-based ELISA developed is a rapid, sensitive and specific detection method and can be used for quantitative characterization of C. perfringens alpha-toxin.
KW - Clostridium perfringens
KW - ELISA
KW - alpha-toxin
KW - monoclonal antibodies
KW - phospholipase C
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U2 - 10.4315/0362-028X-59.6.621
DO - 10.4315/0362-028X-59.6.621
M3 - Article
C2 - 31159017
AN - SCOPUS:0029781913
VL - 59
SP - 621
EP - 625
JO - J.MILK.FOOD TECHNOL.
JF - J.MILK.FOOD TECHNOL.
SN - 0362-028X
IS - 6
ER -