The Menlo Roundtable

Erythropoietin Manufacturing and Purification with Recombinant DNA Plasmids to Treat Anemias

Protein manufacturing and purification with recombinant plasmids is a way to express a desired protein using genetically modified bacteria cells.

This study aims to manufacture and purify recombinant human erythropoietin (rHuEpo), a hormone that stimulates red blood cell production in the bone marrow. This study begins with the construction of a recombinant plasmid made from a gene of interest and a backbone plasmid using Gibson Assembly. After this step, the bacteria is transformed to allow them to obtain the recombinant plasmid. After this, the transformed bacteria are placed on LB agar plates to multiply and produce the protein. After this, erythropoietin is expressed and the protein is purified. This process begins with the lysing of the transformed bacteria cells, and the use of nickel beads to isolate the desired erythropoietin. After the purification is verified by SDS page gel, it will be confirmed that the protein was isolated.

In this study we propose the utilization of synthetic EPO’s blood cell production properties to treat Sickle Cell Disease patients, who struggle with low oxygen content in their blood due to anemia. The process by which EPO leads to the production of red blood cells begins with the production of EPO by interstitial fibroblasts in the kidneys. EPO, which is soluble in water, will then circulate in the body and only interact with cells that have EPO receptors. When EPO gets to the bone marrow, it will interact with red blood cell precursors which have EPO receptors to prevent them from undergoing apoptosis, and instead cause them to become red blood cells. 

Photo: Photo by National Cancer Institute / Unsplash