Development of a Sandwich ELISA Test for Arginase Measurement Based on Monoclonal Antibodies

Abstract

Human arginase was purified from liver and two monoclonal antibodies (MAbs), HA1 and HA2, were produced by fusion of spleen cells from an arginase-immunized BALB/c mouse and the NS-1 myeloma cell line. Both MAbs were of the IgG3 subclass and contained the kappa light chain. HA1 inhibited arginase activity, suggesting that it binds to the arginase catalytic site. HA1 and a horseradish peroxidase-conjugated polyclonal rabbit anti-human arginase antibody were used to develop a sandwich enzyme-linked immunoadsorbent assay (ELISA) for the quantification of human arginase, which can be used in the 1 to 300 ng/mL range. Because of its sensitivity and specificity, this MAb can be successfully applied to the ELISA quantification of arginase in serum and culture supernatants.

Gold Nanoparticle-Based Photoluminescent Nanoswitch Controlled by Host–Guest Recognition and Enzymatic Hydrolysis for Arginase Activity Assay

“…Arginase is most abundantly expressed in mammalian liver, besides being found in nonhepatic tissues, such as red blood cells, kidney, and lactating mammary glands. 43 To assess the performance of this newly constructed method, we apply it to detect arginase activity in real biological samples. The standard addition method is employed to monitor the recoveries (related to the proposed method) in the rat liver sample.…”