title: CIEF method optimization: development of robust and reproducible protein reagent characterization in the clinical immunodiagnostic industry.
abstract: Several method parameters have been refined for application of CIEF methods to provide optimal capillary robustness and performance longevity while maintaining desired analytical output for the ever increasing characterization scrutiny of protein reagents used in clinical assay formulations. Demonstrated here are significant modifications to the existing protocols in order to attain a robust, reproducible method that achieves as much as a 20-fold increase in the number of consecutive runs before capillary degradation. Not only is it a concern for the rudimentary analysis of acidic and basic components of the isoform profile for monoclonal antibodies, but a comprehensive identification of each individual isoform to obtain a characteristic fingerprint is necessary for minor distinguishable properties between multiple proteins in unambiguous identification. In order to maintain the integrity of these modifications, extensive studies were conducted on an implemented system suitability standard protein with specifically defined parameters indicating either sufficient or poor separation performance.
authors: Abbott Laboratories, Abbott Diagnostics Division, Research and Development, IL 60064, USA. ryan.bonn@abbott.com
authors: Ryan
authors: R
authors: Bonn
authors: Sushma
authors: S
authors: Rampal
authors: Tracey
authors: T
authors: Rae
authors: Jeffrey
authors: J
authors: Fishpaugh
mesh_terms: D000911:Antibodies, Monoclonal / Q000032:analysis*; D019075:Electrophoresis, Capillary / Q000379:methods; D007159:Immunologic Tests / Q000379:methods; D007525:Isoelectric Focusing / Q000379:methods* / Q000592:standards; D020033:Protein Isoforms / Q000032:analysis*; D015203:Reproducibility of Results
chemical_list: D000911:Antibodies, Monoclonal; D020033:Protein Isoforms