title: Asymmetrical flow field-flow fractionation for the analysis of PEG-asparaginase.
abstract: Monomethoxypolyethylene glycol L-asparaginase (PEG-ASNASE) is the PEGylated version of the enzyme L-asparaginase (ASNASE). Both are used for remission induction in acute lymphoblastic leukemia (ALL) and non-Hodgkin's lymphoma (NHL). The treatment control is generally carried out by performing activity assays, though methods to determine the actual enzyme rather than its activity are rare. Using asymmetrical flow field-flow fractionation (AF4) offered the chance to develop a method capable of simultaneously measuring PEG-ASNASE and PEG. A method validation was performed in accordance with FDA guidelines for PEG-ASNASE from non-biological solutions. The method unfolded a linearity of 15-750 U/mL with coefficients of correlation of r(2)>0.99. The coefficients of variation (CV) for within-run and between-run variability were 1.18-10.15% and 2.43-8.73%, respectively. Furthermore, the method was used to perform stability tests of the product Oncaspar® (PEG-ASNASE) and estimation of the molecular weight by multi-angle light scattering (MALS) of stressed samples to correlate them with the corresponding activity. The findings indicate that Oncaspar® stock solution should not be stored any longer than 24 h at room temperature and cannot be frozen in pure aqueous media. The validated method might be useful for the pharmaceutical industry and its quality control of PEG-ASNASE production.
authors.affiliation: Institute of Pharmaceutical Technology and Biopharmacy, University of Muenster, Corrensstr. 48, Muenster 48149, Germany.
authors.forename: C
authors.initials: C
authors.lastname: John
authors.affiliation: Department of Pharmaceutical and Medicinal Chemistry - Clinical Pharmacy, University of Muenster, Corrensstr. 48, 48149 Muenster, Germany.
authors.forename: T
authors.initials: T
authors.lastname: Herz
authors.affiliation: Department of Pediatric Hematology and Oncology, University Children's Hospital Muenster, Muenster, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Boos
authors.affiliation: Institute of Pharmaceutical Technology and Biopharmacy, University of Muenster, Corrensstr. 48, Muenster 48149, Germany.
authors.forename: K
authors.initials: K
authors.lastname: Langer
authors.affiliation: Department of Pharmaceutical and Medicinal Chemistry - Clinical Pharmacy, University of Muenster, Corrensstr. 48, 48149 Muenster, Germany. Electronic address: georg.hempel@uni-muenster.de.
authors.forename: G
authors.initials: G
authors.lastname: Hempel
mesh_terms: D001215:Asparaginase / Q000032:analysis* / Q000302:isolation & purification*; D031521:Fractionation, Field Flow / Q000379:methods*; D016014:Linear Models; D011092:Polyethylene Glycols / Q000032:analysis* / Q000302:isolation & purification*; D015203:Reproducibility of Results; D014867:Water / Q000737:chemistry
chemical_list: D014867:Water; D011092:Polyethylene Glycols; C042705:pegaspargase; D001215:Asparaginase
keywords: Asymmetrical flow field-flow fractionation; PEG-asparaginase; Stability