Reduction of matrix interferences by the combination of chaotropic salt and DMSO in a broadly applicable target-based ELISA for pharmacokinetic studies of therapeutic monoclonal antibodies.
Use of a synergistic effect of DMSO together with a chaotropic salt (NaSCN or MgCl2) allowed to drastically reduce matrix interferences in an ELISA for therapeutic monoclonal antibodies. Optimum combinations were found to be 0.4 M NaSCN together with 10.0% DMSO, and 1.0 M MgCl2 with 15.0% DMSO. At this optimum combination, quality controls spiked with mAb at 50.0 ng/ml in eighteen individual human sera and plasmas were quantified with an overall accuracy of 102.0%. All of these QCs fulfilled the acceptance criteria of 80.0-120.0% accuracy and precision below 20.0%. The assay was also successfully applied to the quantification of two other mAbs in human serum. Furthermore, the use of the assay was extended to pre-clinical species (cynomolgus monkey and rat serum). Here, the performed validation experiments confirmed the utility of the assay and demonstrated that the assay allowed quantification of mAb from 50.0 ng/ml to 100.0 microg/ml in cynomolgus monkey serum. The method has then been applied to a pharmacokinetic study in cynomolgus monkeys. In summary, this work demonstrates the efficacy of the combination of a chaotropic salt with DMSO to minimize matrix interferences in an ELISA. The robustness thus obtained allowed the successful establishment of a cost effective, target-based ELISA format for use in pharmacokinetic studies, that is easily applicable for the quantification of mAbs in various matrices such as human, cynomolgus monkey or rat serum and plasma.
Doucet
Julie
J
Department of Marker Localization and Assays, Novartis Institute for Biomedical Research, Klybeckstrasse 141, CH-4057 Basel, Switzerland. julie.doucet@novartis.com
Canadi
Jasna
J
Kalis
Christoph
C
Valentin
Marie-Anne
MA
Marrony
Séverine
S
Deckert-Salva
Fabienne
F
Legay
François
F
Avrameas
Alexandre
A
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry*; D001710:Biotin / Q000737:chemistry; D002626:Chemistry, Pharmaceutical / Q000379:methods*; D004121:Dimethyl Sulfoxide / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay / Q000295:instrumentation* / Q000379:methods*; D006801:Humans; D008252:Macaca fascicularis; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry; D015203:Reproducibility of Results; D012492:Salts / Q000737:chemistry*; D012997:Solvents / Q000737:chemistry; D013678:Technology, Pharmaceutical / Q000379:methods
D000911:Antibodies, Monoclonal; D011994:Recombinant Proteins; D012492:Salts; D012997:Solvents; D001710:Biotin; D004121:Dimethyl Sulfoxide