A new evaluation technique for the detection of impurities in purified proteins via CE with native UV-LIF.
An analytical methodology for quality control analyses of IgG and their impurities is presented using a new UV-LIF (266 nm) detector inside the cassette of a CE instrument and its performance was evaluated. The observed sensitivity was very close to that obtained by silver staining of slab gels (LOD of 25 ng/mL), while the sensitivity of the analysis is 80 times better than with CE/UV absorption (214 nm). Examples of the analysis of pharmaceutical and other commercial IgGs are provided and the kinetics of the reduction of IgG by beta-mercaptoethanol is reported, demonstrating the ease of performing the analysis.
Rodat
Audrey
A
Laboratoire des IMRCP, Université de Toulouse, Université Paul Sabatier, Toulouse, France.
Couderc
Bettina
B
Feurer
Bernard
B
Couderc
François
F
D000818:Animals; D004340:Drug Contamination; D019075:Electrophoresis, Capillary / Q000379:methods*; D007074:Immunoglobulin G / Q000737:chemistry; D004364:Pharmaceutical Preparations / Q000737:chemistry*; D011489:Protein Denaturation; D011817:Rabbits; D051381:Rats; D012680:Sensitivity and Specificity; D016622:Silver Staining; D013050:Spectrometry, Fluorescence / Q000379:methods*
D007074:Immunoglobulin G; D004364:Pharmaceutical Preparations