title: Direct determination of residual Pluronic F-68 in in-process samples from monoclonal antibody preparations by high performance liquid chromatography.
abstract: A simple and sensitive high performance liquid chromatography (HPLC) method was developed to determine residual Pluronic F-68 (PF-68) in in-process samples of monoclonal antibody (MAb) preparations. The method permits the direct injection of proteinaceous samples after simple sample dilution and is able to quantitate as low as 50 mg/L of PF-68 in the presence of up to approximately 30 g/L of protein. The PF-68 molecule was separated on a restricted access reversed phase column using a step gradient and then measured by an evaporative light scattering detector (ELSD). The method was successfully applied to demonstrate PF-68 clearance in MAb purification processes. A modified colorimetric method using liquid-liquid extraction and cobalt thiocyanate to derivatize PF-68 is also described. The results obtained by both the HPLC and colorimetric methods were compared. In addition to its ease of use and simplicity, the HPLC method had better accuracy and higher throughput than the colorimetric method.
authors.affiliation: Analytical & Formulation Sciences, Amgen Inc., One Amgen Center Drive, Thousand Oaks, CA 91320, USA.
authors.forename: Hyo Helen
authors.initials: HH
authors.lastname: Chung
authors.forename: Chengfeng
authors.initials: C
authors.lastname: Zhou
authors.forename: Hui Koon
authors.initials: HK
authors.lastname: Khor
authors.forename: Jinshu
authors.initials: J
authors.lastname: Qiu
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry*; D005591:Chemical Fractionation; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003124:Colorimetry; D004340:Drug Contamination; D020442:Poloxamer / Q000032:analysis*; D012044:Regression Analysis; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D000911:Antibodies, Monoclonal; D020442:Poloxamer