Determination of haptoglobin in bovine serum using polyclonal and monoclonal anti-human haptoglobin antibodies

Abstract

Two ELISA procedures to determine haptoglobin (Hp) in bovine serum were developed. Equine haemoglobin was used as the solid phase. Self-developed goat polyclonal antibody (variant I) and monoclonal antibody (variant II) raised against human Hp were used. The results were compared with the guaiacol method. High correlation was found (r = 0.96 and r = 0.90, respectively) based on the results of 548 bovine serum samples, of which 357 were from clinically healthy cows and 191 from cows and calves monitored during treatment for the most common diseases. The Hp detection limit of ELISA using polyclonal Ab was 0.1 mg/l and using MoAb 0.21 mg/l. The addition of 2% PEG 6000 at the antibody-binding steps enabled major shortening of the incubation periods. The relatively short time, low cost of reagents, and high correlation with the reference method support the use of these ELISA variants in bovine diagnostics.