Expression of human serum albumin in Nicotiana benthamiana

Abstract

Backgrounds: Human serum albumin (HSA), which is mostly produced from human plasma, is limited by supply limits and
safety issues related to potential contamination with blood-borne infections. Thus, choosing a suitable expression system
for recombinant HSA synthesis becomes essential for addressing clinical and cell culture application requirements.
Objective(s): This study explored the use of Nicotiana benthamiana as an alternative platform for producing recombinant
HSA.
Methods: The N. benthamiana plants as an expression platform were used for transient expression of HSA. The agrobacterium
solution containing HSA expression vector was infiltrated into the leaves. A single step Ni-NTA affinity chromatography
method was used to purify the plant-produced HSA. The liquid chromatography-mass spectrometry (LC/MS) was used to
verify the correctness of plant-produced HSA.
Results: We successfully expressed and purified HSA from the leaves of infiltrated N. benthamiana. The final HSA yield
was estimated to be around 30 mg HSA/kg of N. benthamiana lea fresh weight. We identified the intact mass and peptide
mapping of plant-produced HSA, the results suggested to be around 69.07 kDa and a total of 93.52% of the HSA sequence
was found
Conclusion: These results suggest a promising beginning for the advancement of plant-based HSA in the area of plant
molecular farming. Further application needs to be investigated.