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Capillary gel electrophoresis for the quantification and purity determination of recombinant proteins in inclusion bodies
Author(s) -
Espinosade la Garza Carlos E.,
PerdomoAbúndez Francisco C.,
CamposGarcía Víctor R.,
Pérez Néstor O.,
FloresOrtiz Luis F.,
MedinaRivero Emilio
Publication year - 2013
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201300232
Subject(s) - repeatability , recombinant dna , chromatography , chemistry , capillary electrophoresis , coefficient of variation , resolution (logic) , biochemistry , gene , artificial intelligence , computer science
In this work, a high‐resolution CGE method for quantification and purity determination of recombinant proteins was developed, involving a single‐component inclusion bodies ( IB s) solubilization solution. Different recombinant proteins expressed as IB s were used to show method capabilities, using recombinant interferon‐β 1b as the model protein for method validation. Method linearity was verified in the range from 0.05 to 0.40 mg/mL and a determination coefficient ( r 2 ) of 0.99 was obtained. The LOQ s and LOD s were 0.018 and 0.006 mg/mL, respectively. RSD for protein content repeatability test was 2.29%. In addition, RSD for protein purity repeatability test was 4.24%. Method accuracy was higher than 90%. Specificity was confirmed, as the method was able to separate recombinant interferon‐β 1b monomer from other aggregates and impurities. Sample content and purity was demonstrated to be stable for up to 48 h. Overall, this method is suitable for the analysis of recombinant proteins in IB s according to the attributes established on the I nternational C onference for H armonization guidelines.