title: Primary structure control of recombinant proteins using high-performance liquid chromatography, mass spectrometry and microsequencing.
abstract: The conformity of two recombinant proteins (a von Willbrand factor fragment and human serum albumin, consisting of respectively 289 and 585 amino acids) has been examined by HPLC combined with mass spectrometry and microsequencing, on both intact material and fragment peptides obtained by proteolytic cleavage. These studies confirmed that the primary structure of the recombinant proteins corresponds to that predicted from their gene, particularly the integrity of their N and C termini, and, in the case of albumin, the agreement between the observed disulfide bond pattern and the published model. Furthermore, the structure of an albumin-related compound could be elucidated. Application of LC-MS for batch-to-batch quality control is also under discussion.
authors.affiliation: Rhône-Poulenc Rorer S.A., Centre de Recherches de Vitry-Alfortville, Vitry sur Seine, France.
authors.forename: F F
authors.initials: FF
authors.lastname: Clerc
authors.forename: B
authors.initials: B
authors.lastname: Monégier
authors.forename: D
authors.initials: D
authors.lastname: Faucher
authors.forename: F
authors.initials: F
authors.lastname: Cuiné
authors.forename: C
authors.initials: C
authors.lastname: Pourcet
authors.forename: J C
authors.initials: JC
authors.lastname: Holt
authors.forename: S Y
authors.initials: SY
authors.lastname: Tang
authors.forename: A
authors.initials: A
authors.lastname: Van Dorsselaer
authors.forename: J
authors.initials: J
authors.lastname: Becquart
authors.forename: M
authors.initials: M
authors.lastname: Vuilhorgne
mesh_terms: D000595:Amino Acid Sequence; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002851:Chromatography, High Pressure Liquid; D003488:Cyanogen Bromide; D006801:Humans; D006868:Hydrolysis; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011994:Recombinant Proteins / Q000032:analysis*; D017421:Sequence Analysis; D012697:Serine Endopeptidases; D012709:Serum Albumin / Q000032:analysis; D014357:Trypsin; D014841:von Willebrand Factor / Q000032:analysis
chemical_list: D011994:Recombinant Proteins; D012709:Serum Albumin; D014841:von Willebrand Factor; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase; D014357:Trypsin; D003488:Cyanogen Bromide