Radioimmunoassay of Morphine

Synthesis of Immunogen Morphine is first converted to 3-o-carboxymethyl-morphine by reacting the free base with sodium-p-chloroacetate in absolute ethanol  

Antiserum Production The immunogen, carboxymethylmorphine-bovine-serum-albumin, is emulsified with equal volume of complete Freund s adjuvant. Initial immunization doses are injected into the New Zealand albino rabbits and later on this followed up with booster injections after a period of 6 weeks. The antiserum titer is determined with each booster dose injection and is duly harvested when the titre value is maximum. This is diluted suitably and employed in the radioimmunoassay.  

RIA-Procedure The various steps followed are as stated below, namely  

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M Koida, M Takahashi, H Kaneto. The morphine 3-glucuronide directed antibody Its immunological specificity and possible use for radioimmunoassay of morphine in urine. Jpn J Pharmacol 24 707, 1974. 

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