Proteomics

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Determination of Supplier-To-Supplier and Lot-To-Lot Variability in Glycation of Recombinant Human Serum Albumin Expressed in Oryza sativa

ABSTRACT: In studies presented here, recombinant human serum albumins (rHSA) produced in Oryza sativa (Asian rice) (OsrHSA) from a number of suppliers have been extensively characterized and compared to plasma-derived HSA (pHSA) and rHSA expressed in yeast (Pichia pastoris and Saccharomyces cerevisiae). Modifications of the samples were identified by liquid chromatography-mass spectrometry (LC-MS) with samples further assessed with size exclusion chromatography (SEC), reversed-phase high-performance liquid chromatography (RP-HPLC), capillary electrophoresis (CE) as well as additionally biophysical techniques. LC-MS analysis identified a greater number of hexose-glycated arginine and lysine residues on OsrHSA compared to pHSA or rHSA expressed in yeast. Lot-to-lot variability of OsrHSA glycation from one manufactures supplied products were further identified. The extensive glycation of OsrHSA from multiple suppliers may have further implications for the use of OsrHSA as a therapeutic product.

INSTRUMENT(S): Synapt MS

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Marybeth Creskey  

LAB HEAD: Michael J.W. Johnston

PROVIDER: PXD001248 | Pride | 2014-09-12

REPOSITORIES: Pride

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20140130_OsrHSA-ams.mgf Mgf
20140130_OsrHSA-ams.pride.mgf.gz Mgf
20140130_OsrHSA-ams_Chym01.raw.zip Raw
20140130_OsrHSA-ams_Chym02.raw.zip Raw
20140130_OsrHSA-ams_Chym03.raw.zip Raw
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Publications

Determination of supplier-to-supplier and lot-to-lot variability in glycation of recombinant human serum albumin expressed in Oryza sativa.

Frahm Grant E GE   Smith Daryl G S DG   Kane Anita A   Lorbetskie Barry B   Cyr Terry D TD   Girard Michel M   Johnston Michael J W MJ  

PloS one 20141009 10

The use of different expression systems to produce the same recombinant human protein can result in expression-dependent chemical modifications (CMs) leading to variability of structure, stability and immunogenicity. Of particular interest are recombinant human proteins expressed in plant-based systems, which have shown particularly high CM variability. In studies presented here, recombinant human serum albumins (rHSA) produced in Oryza sativa (Asian rice) (OsrHSA) from a number of suppliers hav  ...[more]

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