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Quantitative analysis of hIgG1 in monkey serum by LC-MS/MS using mass spectrometric immunoassay
Christian Lanshoeft1,2, Olivier Heudi1, Sarah Cianférani2
1Novartis Institutes for Biomedical Research, Drug Metabolism & Pharmacokinetics, Novartis Campus, Fabrikstrasse 14, 4056 Basel, Switzerland.
Aim:
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.
Conclusion:
The present assay is suitable for quantitative analysis of hIgG1-based therapeutic proteins in monkey serum at low levels.
