Question

Therapeutic proteins are expressed in mammalian Chinese Hamster Ovary Cells (CHO), which are grown in rich media broth that often is supplemented with proteins. In this cell culture broth, a receptor containing an Fc group (known as an Fc-fusion protein) was expressed at a titer of 1g receptor/L of cell culture broth. This particular receptor is used to treat Rheumatoid Arthritis by acting as a decoy receptor for Tumor Necrosis Factor alpha and decreasing its binding to its native receptor in vivo. The decoy receptor has a pI range of 4.2-5.2 and a molecular weight of 150000 Daltons. During the GMP manufacture of this decoy receptor, a cell culture supplemental peptide was used for the cell culture growth at 1g/L of cell culture broth. The peptide has a molecular weight of 7649 Daltons.  

1. Describe how you can purify the decoy receptor away from the peptide.

Answer 1

A decoy receptor is a receptor which is in a position to understand as well as bind certain growth factors or maybe cytokines effectively, but isn't structurally in the position to mean and trigger the planned receptor complicated. It functions as an inhibitor, binding a ligand and also preventing it from binding to the normal receptor of its. Decoy receptors participate in the same techniques of signal inhibition and can also be plentiful in malignant tissues, which makes up a tremendous subject of cancer research.Receptors can be purified directly utilizing ligands as affinity reagents. Fractionation of receptors on columns to which ligands have been immobilized offers an effective way for purification.