PROTEIN MANUFACTURE IN TRANSGENIC LUZERN PLANTS

摘要

This invention is directed to characterizing a host system suitable for the production of functional transgenic proteins, such as anti-human IgG, for use in applications requiring Government regulatory approval. It is well known that regulatory agencies required stable, consistent master cell banks and master cel l lines for the production of transgenic proteins in order to ensure sufficient material for appropriate characterization, clinical trials, and potential sales. Current plan t production systems require the establishment of seed banks for this purpose. However, there are many draw backs related to such a system for the production o f a continuous reliable transgenic protein source. An aspect of this invention is di rected to characterizing a plant production system suitable for transgenic proteins tha t meet the stringent regulatory requirements. Another aspect of this invention exemplif ies the production and characterization of an anti-human IgG for use as a blood grou ping reagents, through the expression of corresponding genes in transgenic alfalfa pl ants. 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 infectio n. Transgenic plants expressing the light- and heavy-chain encoding mRNAs were obtained and plants from the Fl progeny (obtained by sexual crossing) were found to express fully assembled C 5-1. Furthermore, the transgenic protein was stable in vivo, as well as during extraction and purification procedures. Purification yielded a unique H2L2 form with a reactivity indistinguishable from hybridoma-derived C 5-1 in standardized serological tests. Results indicate that plant-derived transgenic p roteins, such as mAbs can be used as diagnostic reagents as effectively as hybridoma-deri ved mAbs, and demonstrates the usefulness of the transformed alfalfa system to produ ce large amounts of proteins, including multimeric proteins such as mAbs.