Monoclonal Antibody Supernatant and Ascites Fluid Production

2000 ◽  
Vol 40 (1) ◽  
Author(s):  
Wayne M. Yokoyama
2014 ◽  
Vol 2014 ◽  
pp. 1-8
Author(s):  
Shannon T. Marko ◽  
Stephen F. Little ◽  
Carrie G. Benton ◽  
Richard Kelly ◽  
Amy E. Field ◽  
...  

Monoclonal antibody (mAb) ascites fluid production in mice is a well described method of antibody production, although ethical questions regarding the pain and distress of the animals utilized in this process have been raised. In this study, mice were injected with pristane to initiate granuloma formation, followed by an injection of murine hybridoma PA 2II 2F9-1-1 (2F9) to produce IgG1 subclass mAb directed against protective antigen (PA) protein of Bacillus anthracis. Upon the recognition of pain or distress, characterized by well accepted clinical signs, analgesics were administered by treatment group. The control group (A) received saline, group (B) received meloxicam, group (C) received buprenorphine, and group (D) received both meloxicam and buprenorphine. Analgesics were administered by group for a total of 36–48 hours prior to the second ascites fluid collection. There was no statistical difference in the antibody titer or functionality (P>0.05) between treatment groups at the first or the second collection time points. As reported here, analgesics may be administered upon recognition of pain in mice used for mAb ascites fluid production without affecting antibody concentration or quality and may warrant further evaluation as a refinement in other hybridoma cell lines.


Author(s):  
Berma M. Kinsey ◽  
Roger M. Macklis ◽  
James M. Ferrara ◽  
Warren W. Layne ◽  
Steven J. Burakoff ◽  
...  

1996 ◽  
Vol 59 (6) ◽  
pp. 621-625 ◽  
Author(s):  
ZONGLIN L. LIU ◽  
HANS P. BLASCHEK

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.


1991 ◽  
Vol 95 (4) ◽  
pp. 454-459 ◽  
Author(s):  
Franco Bonetti ◽  
Maurizio Pea ◽  
Guido Martignoni ◽  
Aldo Mombello ◽  
Romano Colombari ◽  
...  

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