The use of different expression systems to produce the same recombinant @entity1 protein can result in expression-dependent chemical modifications (CMs) leading to variability of structure, stability and immunogenicity. Of particular interest are recombinant @entity1 proteins expressed in plant-based systems, which have shown particularly high CM variability. In studies presented here, recombinant human serum albumins (rHSA) produced in @entity579 (Asian @entity579 ) (OsrHSA) from a number of suppliers have been extensively characterized and compared to plasma-derived HSA (pHSA) and rHSA expressed in @entity375 ( @entity10056 and @entity375 ). The heterogeneity of each sample was evaluated using size exclusion chromatography (SEC), reversed-phase high-performance liquid chromatography (RP-HPLC) and capillary electrophoresis (CE). Modifications of the samples were identified by liquid chromatography-mass spectrometry (LC-MS). The secondary and tertiary structure of the albumin samples were assessed with far U/V circular dichroism spectropolarimetry (far U/V CD) and fluorescence spectroscopy, respectively. Far U/V CD and fluorescence analyses were also used to assess thermal stability and drug binding. High molecular weight aggregates in OsrHSA samples were detected with @entity16267 and supplier-to-supplier variability and, more critically, lot-to-lot variability in one manufactures supplied products were identified. LC-MS analysis identified a greater number of @entity12015 -glycated @entity369 and @entity376 residues on OsrHSA compared to pHSA or rHSA expressed in @entity375 . This analysis also showed supplier-to-supplier and lot-to-lot variability in the degree of glycation at specific @entity376 and @entity369 residues for OsrHSA. Both the number of glycated residues and the degree of glycation correlated positively with the quantity of non-monomeric species and the chromatographic profiles of the samples. Tertiary structural changes were observed for most OsrHSA samples which correlated well with the degree of @entity369 / @entity376 glycation. The extensive glycation of OsrHSA from multiple suppliers may have further implications for the use of OsrHSA as a therapeutic product.

Determination of Supplier-to-Supplier and Lot-to-Lot Variability in Glycation of Recombinant Human Serum Albumin Expressed in XXXX .

@entity1 :: ('9606', 'Species') :: ['human'] @entity10056 :: ('4922', 'Species') :: ['Pichia pastoris'] @entity16267 :: ('CHEBI:30003', 'Chemical') :: ['SEC'] @entity12015 :: ('CHEBI:18133', 'Chemical') :: ['hexose'] @entity376 :: ('CHEBI:25094', 'Chemical') :: ['lysine'] @entity375 :: ('4932', 'Species') :: ['yeast', 'Saccharomyces cerevisiae'] @entity579 :: ('4530', 'Species') :: ['Oryza sativa', 'rice'] @entity369 :: ('CHEBI:29016', 'Chemical') :: ['arginine']
@entity579 :: (4530,Species) :: ['Oryza sativa']
