Problems to Mab production

Gianpaolo Grassi ggrassi at icgeb.trieste.it
Wed Apr 17 12:53:32 EST 1996

Dear Netters,
I am in trouble for a big problem to produce Mab against 
tetrahydrocannabinol THC (delta-9-THC). I am using 3 different carrier 
proteins, BSA, KLH, porcine Thyroglobuline; 2 aptenes delta-9 and delta 8 
THC; 2 conjugation methods, emisuccinate and butiletere method; 4 adjuvants, 
FCA, Ribi's , Titermax, Gerbu; 2 mouse strains, BALB/c and C57;different 
immunisation schedules, intrasplenic injections, adsorbtion of antigen on 
nitrocellulose, transplantation of spleen cells to irradiated mice, some 
attempts to in-vitro immunisation and some other trials but until now I 
have no Mab against THC. This Mab is available from private companies so 
it is possible to produce it but I would like to obtain a line for my own 
use. Please let me know some advice or what are the problems with this 
aptene. The mice I have used to fusions (10 for about 3000 clone tested) 
produce policlonal sera with low titre (max 1:1000) and at the first 
screen tests I have some positive clones but after few ELISA control they 
stop to produce IgG. I have cloned all the positive clone immediatelly but no 
good results. I have tested myeloma with PCR for mycoplasma but no 
infection. I am looking forward to obtain some help,
G. Grassi,

E-Mail isci at www.netuno.it
Istituto Sperimentale per le Colture Industriali
Via di Corticella, 133
40129 Bologna Italy  

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