Mass spectrometry study of a transferrin-based protein drug reveals the key role of protein aggregation for successful oral delivery |
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Authors: | Cedric E Bobst Shunhai Wang Wei-Chiang Shen Igor A Kaltashov |
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Institution: | Department of Chemistry, University of Massachusetts, Amherst, MA 01003. |
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Abstract: | A recently designed human growth hormone/transferrin fusion protein (GHT) remains one of the very few examples of a protein capable of eliciting measurable therapeutic response after oral administration. To better understand the underlying factors that resulted in this rare success of nonparenteral protein drug delivery, we analyzed proteolytic stability and receptor binding properties of this protein, the key factors in overcoming the primary barriers to successful oral delivery. Analysis of GHT by a combination of size exclusion chromatography and mass spectrometry revealed that a significant protein population exists in an oligomeric (GHTx) state in addition to the anticipated monomer (GHT1). These states of GHT were evaluated for their survivability in stomach-like conditions, as well as their ability to bind transferrin receptor (TfR). Our results reveal an exceptional stability of GHTx, as well as the preserved ability to bind TfR, a critical first step in crossing the epithelial-intestinal barrier through receptor-mediated transcytosis. |
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Keywords: | protein therapeutic electrospray ionization native electrospray ionization MS protein/receptor interaction recombinant fusion protein |
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