Patent ID: 6740526
Filing Date: 2004-05-25
Classification: C07K,C12N,C12P

Abstract:
A method for transiently transforming plant cells or plant tissue for the large scale production of recombinant polypeptide comprising:i) providing a plant tissue sample to a bioreactor or cultivating plant cells or plant tissue in liquid medium in a bioreactor under conditions suitable for growth of the cells or tissue, ii) inoculating the plant cells or plant tissue with a culture of Agrobacteria when suitable growth of the cells or tissues is obtained, the Agrobacteria containing a vector comprising a nucleotide sequence encoding the recombinant polypeptide; iii) culturing the plant cells or plant tissue and the Agrobacterium under conditions suitable for transfer of the nucleotide sequence to the plant cells or the plant tissue to thereby produce transiently transformed plant cells or plant tissue, iv) growing the transiently transformed plant cells or plant tissue in liquid medium for about one to about four days under conditions that enable the transiently transformed plant cells or tissue to transiently express the recombinant polypeptide; and v) recovering the recombinant polypeptide from the transiently transformed cells or tissue of step (iv), wherein the conditions are monitored during step (i), (iii), and/or (iv) by measuring optical density, pH, temperature, nutrient levels, oxygen, conductivity, refractive index, osmolarity, calcium level of the medium, protein expression level, or a combination thereof; and wherein the Agrobacterium is an auxotroph deficient in its ability to metabolize amino acids, vitamins, and/or nucleic acid precursors, wherein said auxotroph has impaired ability to grow in a plant root or cell culture.