Penggunaan metode radioassay teknik fase padat dalam reaksi fiksasi a--Kobratoksin terhadap reseptor koligemik

USING OF SOLID PHASE RADIOASSAY MEnlOD IN THE FIXAllON REACTION OF aCOBRA
TOXIN TO CHOliNERGIC RESEPTOR. a-cobratoxin is a protein tl1at is in the snake venom of
Naja naja kaouthia. If, it enter into the body, would be fixed specifically to the cholinergic receptor, causing
muscle and breathing paralyze. Determination of functional groups in this fixation could be accomplish by
filtration radioassay method with labelled radioactive a-cobratoxin. Beside that, the functional groups in this
fixation could be determined by solidPl1ase raaJoassay method. The determination of optimum condition of acobratoxin
fixation to cholinergic receptor with labeled tritium a-cobratoxin (3H-a-cobratoxin) using this method
has been carried out. Optimum condition of fixation was reached at 20 J:tg of cholinergic receptor coated on the
ELISA plat with ~-a-cobratoxin activity range 5.0 104 to 7.5 104 cpm and 4 hours of minimum time incubation
at room temperature. Competitive reaction betweell ~-a-cobratoxin and a-cobratoxin to the cholinergic receptor
gave 6.50 10-9 M of the 50% of inhibition competitive value (ICso) Evaluation of this method using (N.-acetyllysin-23)a-cobratoxin
derivative showed tl1athe compound was inhibited in the fixation to cholinergic receptor
with IC5o = 9.50 10-9 M. From this experiment was obtained that this method could be used to determine the
active group of the toxin which function in the fixation reaction to cholinergic receptor.