Discrimination among IgG1-kappa monoclonal antibodies produced by two cell lines using charge state distributions in nanoESI-TOF mass spectra.
Charge state distributions (CSDs) of proteins in nanoESI mass spectra are affected by the instrumental settings and experimental conditions, in addition to the conformations of the proteins in the analyzed solutions. In the presented study, instrumental and experimental parameters--the desolvation gas flow rate, temperature, pH, buffer (ammonium acetate), and organic modifier (methanol) concentrations--were optimized according to a reduced central composite face experimental design to maximize the separation of CSDs of monoclonal IgG1-kappa antibodies produced by two production systems (CHO and GS-NS0 cell lines). Principal component analysis and Fisher linear discriminant analysis were then used to reduce the dimensions of the acquired dataset and quantify the separation of the protein classes, respectively. The results show that the IgG1-kappa molecules produced by the two production systems can be clearly distinguished using the described approach, which could be readily applied to other proteins and production systems.
Zamani
Leila
L
Department of Analytical Chemistry, Stockholm University, Stockholm, Sweden.
Lindholm
Jessica
J
Ilag
Leopold L
LL
Jacobsson
Sven P
SP
D000085:Acetates / Q000737:chemistry; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000145:classification; D016466:CHO Cells; D045744:Cell Line, Tumor; D006224:Cricetinae; D003412:Cricetulus; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000276:immunology*; D007145:Immunoglobulin kappa-Chains / Q000276:immunology*; D016014:Linear Models; D000432:Methanol / Q000737:chemistry; D051379:Mice; D018448:Models, Immunological; D025341:Principal Component Analysis; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013696:Temperature
D000085:Acetates; D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D007145:Immunoglobulin kappa-Chains; C018824:ammonium acetate; D000432:Methanol