Protein fusion is a genetic recombination technique in which the DNA of an inactive peptide or protein chain segment to be grafted is recombined with the DNA of a drug and expressed together by engineered cells; no specialized grafting operations are required. In the current technology, the inactive peptide or protein chain segment used for protein fusion is the FC region of albumin and antibody. Biological grafting techniques to extend the half-life of drugs also face the problem of blocking the catalytic site of the enzyme while blocking the site of the drug binding to the therapeutic target, reducing the specific activity of the FC chain segment of the albumin or antibody. In addition, based on recombinant grafting, it is difficult to selectively and rapidly degrade the albumin or FC chain segment to expose the original drug molecule. Fusion protein gene stability, regulated expression and simple preparation, homogeneous products, low impact on protein and peptide drug activity, etc., is a good way to study long-acting peptide and protein drugs.
The target gene is firstly cloned, and the cDNA is used as the template to amplify different target DNA fragments by PCR with suitable primer sequences designed according to the principle of gene sequence complementation. Then, the recombinant plasmid was constructed by in vitro ligation of two DNA fragments with the same end-site by restriction endonuclease and cloned into the high expression plasmid vector. Next, the recombinant expression vector was transfected into host cells and screened and sequenced using selection markers. Finally, the fusion gene is induced and the expressed protein is purified.
Fusion genes can be expressed in prokaryotic or eukaryotic cells. Prokaryotic expression systems are characterized by short time and low cost, and are the main tools in research. Eukaryotic expression systems are characterized by high post-translational processing opportunities and can even be modified into human-derived forms; eukaryotic cells can be easily transfected and are genetically stable and reproducible; products can be secreted and purification is simple and low cost.
Using fusion expression technology, CD Formulation is able to modify marketed cytokines to develop a steady stream of new, superior products. As a leading fusion protein technology services company, we have years of experience in long-acting protein-based drugs. Featuring strict quality control and short service cycles, we offer our clients a customizable one-stop service in order to save your valuable time and budget. Our technical services cover every step of your project from gene synthesis to data reporting: gene synthesis, expression plasmid construction - expression of decoy proteins, purification of proteins, washing and elution.
With the development of genetic engineering technology, many protein drugs have been obtained by means of genetic engineering, including peptides, cytokines, coagulation factors, enzymes, hormones, growth factors, etc. The fusion molecule is produced by coupling the cDNA of the drug protein to the cDNA of the desired fusion protein and attaching the drug protein to the C-terminus or N-terminus of the desired fusion protein.
If you are interested in our technology, please do not hesitate to contact us and we will provide you with a customized solution.