Mass spectrometry study of a transferrin-based protein drug reveals the key role of protein aggregation for successful oral delivery
Mass spectrometry study of a transferrin-based protein drug reveals the key role of protein aggregation for successful oral delivery
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DOI：
10.1073/pnas.1206924109
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发表时间：
2012-08-21
影响因子：
11.1
通讯作者：
Kaltashov, Igor A.
中科院分区：
文献类型：
--
作者：
Bobst, Cedric E.;Wang, Shunhai;Kaltashov, Igor A.
最近设计的人生长激素/转铁蛋白融合蛋白（GHT）仍然是口服给药后能够引发可测量的治疗反应的蛋白质的极少数实例之一。为了更好地理解导致这种罕见的非胃肠外蛋白质药物递送成功的潜在因素，我们分析了这种蛋白质的蛋白水解稳定性和受体结合特性，这是克服成功口服递送的主要障碍的关键因素。通过尺寸排阻色谱法和质谱法的组合对GHT的分析显示，除了预期的单体（GHT 1）之外，显著的蛋白质群体以寡聚体（GHTx）状态存在。评价这些状态的GHT在胃样条件下的存活能力以及它们结合转铁蛋白受体（TfR）的能力。我们的研究结果揭示了GHTx的特殊稳定性，以及保留的结合TfR的能力，这是通过受体介导的转胞吞作用穿过上皮-肠屏障的关键第一步。
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.