High Expression and Purification of Recombinant Human Serum Albumin from Pichia pastoris

Area of Science:

• Biotechnology
• Biochemistry
• Molecular Biology

Background:

• Recombinant human serum albumin (rHSA) is a valuable therapeutic protein.
• Efficient and scalable production methods for rHSA are crucial for its widespread application.
• Pichia pastoris is a well-established host for recombinant protein expression.

Purpose of the Study:

• To optimize the expression of recombinant human serum albumin (rHSA) in Pichia pastoris.
• To develop and implement a purification strategy for obtaining highly pure rHSA.
• To achieve a high yield of electrophoretically pure rHSA for potential therapeutic use.

Main Methods:

• Optimization of recombinant protein expression in Pichia pastoris.
• Downstream processing involving hollow-fiber ultrafiltration for initial isolation.
• Multi-step chromatography including Phenyl-Sepharose hydrophobic interaction chromatography and antibody-immunoadsorbent chromatography for purification.

Main Results:

• Achieved an expression level of approximately 150 mg/L of rHSA in Pichia pastoris supernatant.
• Successfully purified rHSA through a combination of ultrafiltration and chromatography.
• Demonstrated electrophoretic purity of the final rHSA product.

Conclusions:

• Pichia pastoris is an effective host for high-level production of recombinant human serum albumin.
• The developed purification protocol effectively isolates and purifies rHSA to a high degree.
• The optimized expression and purification process offers a viable method for producing therapeutic-grade rHSA.