Purification of the food-borne carcinogens 2-amino-3-methylimidazo [4,5-f]quinoline and 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline in heated meat products by immunoaffinity chromatography.

A rapid and simple scheme has been developed for the isolation and purification of two of the major mutagenic heterocyclic amines formed in heated beef products by affinity chromatography using monoclonal antibodies which recognize 2-amino-3-methylimidazo[4,5-f]quinoline (IQ). Two cell lines producing IgG antibodies were established following fusion of Sp2 or P3x.63 myeloma cells with spleen cells of immunized BALB/cby mice. The antigen was bovine gamma globulin haptenized with 2-(3-carboxypropylthio)-3-methylimidazo-[4,5-f]quinoline. The antibodies were immobilized on CNBr-activated Sepharose 4B. IQ and MeIQx formed in heated beef products were partially purified by XAD-2 chromatography and then applied to the affinity columns. Purification by affinity chromatography was adequate for subsequent quantitative analysis by HPLC with UV detection. With this purification scheme as little as 1 g of beef extract or 15 g of fried beef could be assayed for IQ and MeIQx at the part per billion level. Both antibodies had similar affinity constants for IQ (9.3 X 10(6) and 6.7 X 10(6) M-1) and for MeIQx (7.1 X 10(5) and 2.7 X 10(5) M-1) and both were suitable for immunoaffinity purification of IQ from complex mixtures. MAb2 could be used as well to selectively remove MeIQx from meat products after partial purification by XAD-2. MAb1, despite having a 3-fold higher affinity than MAb2 for MeIQx, could not be used for affinity chromatography for this mutagen.

Effect of fibrin-fibrinogen degradation products on human basilar artery preparations.

Experiments were performed to determine the effects of fibrin-fibrinogen degradation products on the human basilar artery in vivo. Citrated plasma and streptokinase were incubated at 37 degrees C to produce a preparation of fibrinogen degradation products. Aliquots of the incubate were obtained at 90 minutes, 48 hours, and 1 and 2 weeks after preparation, and were separated into fractions of different molecular weight (MWt), using an ultrafiltration technique. Each fraction was tested at each of the above times for contractile activity and possible interaction with a threshold concentration of 5-hydroxytryptamine (5-HT) on the human basilar artery. Contractile activity was initially confined to the 90-minute aliquot fraction MWt greater than 100,000, but as the incubation proceeded, such activity was also seen in the lower MWt fraction less than 100,000 greater than 10,000 at all time intervals. This activity was never seen in the fraction MWt less than 10,000 at any time. Enhancement of the 5-HT response was initially confined to the higher molecular weight fractions, but after 48-hour incubation all fractions showed this activity. It is suggested that products of fibrin-fibrinogen degradation may be involved directly or indirectly in influencing the pathophysiological mechanism(s) responsible for cerebral arterial spasm following subarachnoid hemorrhage.