Positive Albumin: Enhancing Recombinant Protein Secretion and Purification in Mammalian Cell Lines
This invention introduces a mutant human serum albumin designed to significantly improve the secretion and purification of recombinant proteins from mammalian cell lines. Its most significant advantage lies in enabling efficient protein purification using cation exchange chromatography, overcoming limitations of current methods.
Description
The invention is a modified human serum albumin featuring 14 mutations of acidic amino acids to lysine, along with a hexahistidine tag after the albumin pro-sequence. This engineered albumin, when genetically fused to a target protein, facilitates its secretion from mammalian cells into the culture media. Unlike traditional methods, the positive charge of the mutant albumin allows for effective binding to cation exchange resin, enabling efficient separation from contaminants and providing purification akin to affinity chromatography. The hexahistidine tag at the N-terminus of the albumin further allows for rapid removal of the albumin and proteases post-separation from the desired recombinant protein. This approach addresses the challenges of protein aggregation and column clogging often encountered with other purification techniques.Applications
- Biopharmaceutical manufacturing for efficient protein production.- Research and development of new protein-based therapeutics.
- Production of diagnostic reagents.
- Biotechnology companies seeking improved protein expression and purification systems.
- Development of high-purity protein products for various applications.
Advantages
- Applicability to most if not all proteins.- Speed of protein purification.
- Ease of removal.
- Substantial cost reduction in the procedure.
- Overcomes limitations of current recombinant protein purification methods in mammalian cell lines.