Quantitative analysis of hIgG1 in monkey serum by LC-MS/MS using mass spectrometric immunoassay.
OBJECTIVE
A sensitive generic LC-MS/MS method for hIgG1 quantification in cynomolgus monkey serum using mass spectrometric immunoassay disposable automation research tips (MSIA-D.A.R.T.'S™) is reported.


RESULTS
The hIgG1 was captured with a biotinylated mouse anti-hIgG antibody (50.0 µg/ml) targeting the fragment crystallizable (Fc) region. Elution from the streptavidin-coated MSIA-D.A.R.T.'s was conducted with 0.4% trifluoroacetic acid in water. The method was selective and linear from 10.0 to 1000 ng/ml using 100 µl of serum. The method was evaluated regarding accuracy, precision, carry-over, dilution, auto-sampler stability and applied for the determination of hIgG1 concentration in monkey serum after intravitreal administration.


CONCLUSIONS
The present assay is suitable for quantitative analysis of hIgG1-based therapeutic proteins in monkey serum at low levels.
Novartis Institutes for Biomedical Research, Drug Metabolism & Pharmacokinetics, Novartis Campus, Fabrikstrasse 14, 4056 Basel, Switzerland.|Laboratoire de Spectrométrie de Masse BioOrganique, Institut Pluridisciplinaire Hubert Curien, CNRS-Université de Strasbourg, UMR 7178, 25 rue Becquerel, 67087 Strasbourg, France.
Christian
C
Lanshoeft
Novartis Institutes for Biomedical Research, Drug Metabolism & Pharmacokinetics, Novartis Campus, Fabrikstrasse 14, 4056 Basel, Switzerland.
Olivier
O
Heudi
Laboratoire de Spectrométrie de Masse BioOrganique, Institut Pluridisciplinaire Hubert Curien, CNRS-Université de Strasbourg, UMR 7178, 25 rue Becquerel, 67087 Strasbourg, France.
Sarah
S
Cianférani
Novartis Institutes for Biomedical Research, Drug Metabolism & Pharmacokinetics, Novartis Campus, Fabrikstrasse 14, 4056 Basel, Switzerland.
Andrew P
AP
Warren
Novartis Institutes for Biomedical Research, Drug Metabolism & Pharmacokinetics, Novartis Campus, Fabrikstrasse 14, 4056 Basel, Switzerland.
Franck
F
Picard
Novartis Institutes for Biomedical Research, Drug Metabolism & Pharmacokinetics, Novartis Campus, Fabrikstrasse 14, 4056 Basel, Switzerland.
Olivier
O
Kretz
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000737:chemistry; D019921:Biotinylation; D002853:Chromatography, Liquid / Q000379:methods; D007118:Immunoassay / Q000379:methods*; D007074:Immunoglobulin G / Q000032:analysis / Q000097:blood*; D057230:Limit of Detection; D008252:Macaca fascicularis / Q000097:blood*; D051379:Mice; D053719:Tandem Mass Spectrometry / Q000379:methods*
D000888:Antibodies, Anti-Idiotypic; D007074:Immunoglobulin G
generic LC–MS/MS assay; generic surrogate peptide; hIgG1; mass spectrometric immunoassay disposable automation research tips; therapeutic proteins