Production and characterization of a specific Rubisco monoclonal antibody,and its use in Rubisco quantification during Zantedeschia aethiopica spathe development

Ribulose 1,5-bisphosphate carboxylase/oxygenase was purified from leaves of Zantedeschia aethiopica and used to immunize female Balb/c mice. Monoclona l antibodies (MAbs) were raised by hybridoma technology using Sp2/0 myeloma cells as fusion partner. A random selected IgG2a subclass MAb was purified from ascitic fluid by affinity chromatography on Protein A-Sepharose CL-4B , with a recovery of 84.3% and it was apparently homogeneous on native PAGE , The monoclonality of the purified MAb was determined by IEF, The MAb was highly specific for Rubisco from leaves of Z. aethiopica as determined by W estern blotting and was used to determine the concentration of Rubisco prot ein by enzyme-linked immunoadsorbent assay (ELISA), at three distinct stage s of Z, aethiopica spathe development and in the leaf. The results suggest de novo synthesis of Rubisco during the spathe regreening, which could expl ain, at least in part, the increase of photosynthetic activity observed dur ing regreening.