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