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A novel immunoenzymatic assay is described for the quantitation of human haptoglobin (Hp). Two binding sites on the Hp molecule, namely for hemoglobin (Hb) and for the specific antibody, are involved in the reaction. Hb adsorbed onto the polystyrene microplate binds Hp present in any biological fluid. The formed Hp-Hb complex is detected with horse-radish peroxidase conjugated with anti-Hp antibody. By means of this ELISA, Hp may be measured in the range of 5 to 150 micrograms/L. Comparison of the Hp-ELISA with two other methods of Hp determination resulted in correlation coefficients of 0.97 to 0.99. Intra- and inter-assay coefficients of variation ranged from 4.7 to 6.7%. Hp levels were measured in urine, cord serum, cerebrospinal fluid, amniotic fluid and saliva.

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Following subacute intoxication of rats with Pb-, Cu-, and Zn-salts (separately or in mixture) for 5 weeks, the chelating agent D-penicillamine was administered for 3 weeks. In the course of the 3-month experiment, lactate dehydrogenase (LDH) was estimated in serum and in cytoplasmic fraction of the kidney. Pb2+ treatment resulted in an increase of LDH activity, Cu2+ in a slight decrease, whereas Zn2+ had no effect, respectively. Mixture of these metals caused a significant rise in the enzymatic activity. Seven weeks after the stoppage of the administration of toxic substances, altered LDH activity, both in serum and in kidney returned to normal. D-penicillamine treatment was found to accelerate a restoration of the enzyme activity. In the experiments in vitro, Cu2+ inhibited significantly the kidney LDH activity, Pb2+ and Zn2+ being 100- and 400-times less efficient, respectively. Cu2+ inhibition was reversed by D-penicillamine, whereas inhibition of LDH by Zn2+ or Pb2+ was irreversible.

Summary:The concentration of haptoglobin in sera of healthy women and patients with non-malignant and malignant ovarian tumours was measured by two methods, i.e. complex formation with haemoglobin and complex formation with concanavalin A. High correlation (r = 0.74) between both the methods was found in the group of healthy women, but correlation coefficients were much lower in the group of non-malignant and malignant tumours (r = 0.42 and 0.37, respectively). Direct determinations of haptoglobin, and calculation of the ratio of haptoglobin bound to concanavalin A to haptoglobin bound to haemoglobin revealed statistically significant differences among the examined groups. Comparison of two methods of haptoglobin quantitation suggest that processes connected with ovarian disorders may alter the glycosylation of haptoglobin oligosaccharide chains.