Multimeric proteins within transgenic alfalfa plants and their use in diagnostic assays

摘要

The characterising of a plant production system suitable for transgenic proteins that meet the stringent U.S. Government regulatory requirements is described. The production and characterisation of an anti-human IgG for use as a blood grouping reagents is exemplified through the expression of corresponding genes in transgenic alfalfa plants. The cDNAs of the heavy and light chains of a human IgG-specific IgG2a(kappa) murine mAb (C5-1) were transferred into alfalfa through Agrobacterium infection. Transgenic plants expressing the light- and heavy-chains encoding mRNAs were obtained and plants from the F1 progeny (obtained by sexual crossing) were found to express fully assembled C5-1. Furthermore, the transgenic protein was stable in vivo, as well as during extraction and purification procedures. Purification yielded a unique H2L2 form with reactivity indistinguishable from hybridoma-derived C5-1 in standardised serological tests. Results indicate that plant-derived transgenic proteins, such as mAbs can be used as diagnostic reagents as effectively as hybridoma-derived mAbs, and demonstrates the usefulness of the transformed alfalfa system to produce large amounts of proteins, including multimeric proteins such as mAbs.