Detection of truncated midkine in Wilms’ tumor by a monoclonal antibody against human recombinant truncated midkine

Abstract

Although the expression of a truncated midkine (tMK) mRNA has been detected in many cancer cells, the tMK protein itself has not yet been identified. The expression, purification and characterization of human recombinant tMK were described in the former report [1]. A mouse hybridoma cell line producing an IgG2b monoclonal antibody (mab) against purified recombinant tMK was established. This anti-tMK mab did not cross react with synthetic full length (or c-half) human midkine. A putative native tMK was identified in G401 cells using this mab, and showed the same apparent Mw as the recombinant tMK in SDS-PAGE. This mab was also used in an immunohistochemical study to evaluate the expression of tMK in Wilms’ tumor cell line, G401 cells, as well as in Wilms’ tumor patient specimens. G401 cells and all Wilms’ tumor patient specimens immunoreacted with this anti-tMK mab. We conclude that Wilms’ tumor cells express tMK and that this mab is useful for the detection of tMK in the Wilms’ tumor.