Quantitative Analysis of Higg1 in Monkey Serum by LC–MS/MS Using Mass Spectrometric Immunoassay

Abstract

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.

Keywords::

• generic LC–MS/MS assay
• generic surrogate peptide
• hIgG1
• mass spectrometric immunoassay disposable automation research tips
• therapeutic proteins

Supplementary Data

Financial & competing interests disclosure

This work was conducted in fulfillment for the degree of PhD (University of Strasbourg) in case of Christian Lanshoeft and was performed at the Novartis facilities in Basel for research purposes only without any relation to other projects being currently under development. The authors have no other relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript apart from those disclosed.

No writing assistance was utilized in the production of this manuscript.

Ethical conduct of research

The authors state that they have obtained appropriate institutional review board approval or have followed the principles outlined in the Declaration of Helsinki for all human or animal experimental investigations. In addition, for investigations involving human subjects, informed consent has been obtained from the participants involved.

Acknowledgements

The authors would like to thank Ravindra Chaudhari (Thermo Fisher Scientific) for providing technical support. Moreover, a special thanks to Birgit Jaitner (DMPK Novartis Basel) for providing the study samples and to Mark Milton (DMPK Novartis Cambridge, MA, USA) for critically reviewing this manuscript.