Creative BioMart has developed an Agrobacterium tumefaciens-mediated gene delivery method to facilitate transient recombinant protein expression. The introduction of DNA fragments through Agrobacterium tumefaciens-mediated T-DNA transfer is a powerful research tool. Most cells within infiltrated areas will receive the T-DNAs. It has the possibility of introducing several different T-DNAs into each cell. So far, we have successfully produced a variety of proteins including but not limited to: allergens, active enzymes, monoclonal antibodies, virus-like particles, human cytokines for gene therapy.
We first start with the vector containing the DNA sequence of the target protein, and then simply put it into a bacterium called Agrobacterium tumefaciens. We use electricity to help push the vector into the bacteria through a process called transformation, and once inside, we use a technology called osmosis to put the bacteria containing the DNA vector in the leaves of plants. We are using a class of carefully studied plants that grow rapidly and are easy to penetrate. Once in the plant, bacteria can transfer the DNA encoding protein to the plant cells, then "transcribe" it into RNA and "translate" it into protein. The process lasts for a few days, and leaves are harvested and proteins are extracted after three to six days.
We use chromatography to separate plant proteins from recombinant proteins. Using the characteristics of target protein, for example, size, charge and hydrophobicity to separate it from unwanted proteins. When there is a special label on the recombinant protein, we usually use affinity chromatography to capture the protein.
The purification of recombinant proteins from plant leaves is very similar to that from other types of cells. Leaves are ground up in a solution that contains all the ingredients needed to keep proteins happy, so buffers and salts are included to replicate the natural conditions in which proteins normally exist. Once these proteins are extracted from plant leaves, they need to be separated from unwanted plant components, such as lignin, cellulose and starch. Filtration and centrifugation are used to separate these large and dense molecules. It is at this point that the recombinant protein is separated from the plant protein in the "protein soup".
In addition to the Agrobacterium Tumefaciens-Mediated method, we also provide other two delivery strategies for clients to choose from. Please click the links below if you are interested.
With our professional research and development team and the latest protein expression and purification resources, we promise to deliver high-quality products for our customers. Customers who need our assistance are more than welcome to contact us or submit an online inquiry. Our professional team will provide you with detailed guidance and solution ASAP.
