Development of quantitative mass spectrometric immunoassay for serum amyloid A

Abstract

Objective: Proteins can exist as multiple proteoforms in vivo that can have important roles in physiological and pathological states.

Methods: We present the development and characterization of mass spectrometric immunoassay (MSIA) for quantitative determination of serum amyloid A (SAA) proteoforms.

Results: Intra- and inter-day precision revealed CVs <10%. Against existing SAA ELISA, the developed MSIA showed good correlation according to the Altman–Bland plot. Individual concentrations of the SAA proteoforms across a cohort of 170 samples revealed 7 diverse SAA polymorphic types and 12 different proteoforms.

Conclusion: The new SAA MSIA enables parallel analysis of SAA polymorphisms and quantification of all expressed SAA proteoforms, in a high-throughput and time-efficient manner.

Keywords:

• Baseline concentration
• inflammation biomarker
• mass spectrometry
• polymorphism
• posttranslational modifications
• proteoforms

Disclosure statement

All authors report no declaration of interest. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institute of Diabetes and Digestive and Kidney Diseases or the National Institutes of Health.

Funding

This work was supported by the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) [grant numbers R01DK082542, R24DK090958] to O.T., C.R.B., R.W.N and D.N., and [grant number K23HL107389] to H.N.Y.