Optimization of a human milk-directed quantitative sIgA ELISA method substantiated by mass spectrometry

Kelly A Dingess; Pauline van Dam; Jing Zhu; Marko Mank; Karen Knipping; Albert J R Heck; Bernd Stahl

doi:10.1007/s00216-021-03468-4

Optimization of a human milk-directed quantitative sIgA ELISA method substantiated by mass spectrometry

Kelly A Dingess, Pauline van Dam, Jing Zhu, Marko Mank, Karen Knipping, Albert J R Heck, Bernd Stahl

Danone Nutricia Research

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

Immunoglobulins are the primary protective products in human milk and are responsible for transferring maternal pathogen memory to the infant, providing protection by binding to recognized pathogens and inhibiting virulence. To better understand potentially protective/anti-infective compounds in human milk, the establishment of human milk-tailored analytical approaches is crucial, as most contemporary analytical methods have been optimized for plasma or serum. One of the most prominent immunoglobulins in human milk is secretory immunoglobulin A (sIgA), which may be relevant for the protection of breastfed infants from harmful pathogens. Advanced sIgA detection methods can help monitor the immune status and development of the mother-infant dyad. We therefore developed an enzyme-linked immunosorbent assay (ELISA) sIgA method for the quantitative analysis of IgA plus secretory component (SC), validated with sIgA standards and substantiated by mass spectrometry (MS)-based proteomics. A very strong correlation was observed between the MS-detected IgA1 and the human milk-specific sIgA ELISA (r = 0.82). Overall, the MS data indicate that the developed human milk sIgA ELISA does not differentiate between sIgA1 and sIgA2 and is, therefore, a reflection of total sIgA. Furthermore, our MS data and the human milk-derived sIgA ELISA data are better correlated than data derived from a standard serum IgA ELISA kit (relative to MS IgA1 r = 0.82 and r = 0.42, respectively). We therefore propose our human milk-specific sIgA ELISA as an ideal quantitative indicator of total sIgA with advantages over current serum IgA ELISA kits.

Original language	English
Pages (from-to)	5037-5049
Number of pages	13
Journal	Analytical and Bioanalytical Chemistry
Volume	413
Issue number	20
Early online date	25 Jun 2021
DOIs	https://doi.org/10.1007/s00216-021-03468-4
Publication status	Published - Aug 2021

Bibliographical note

Funding Information:
Primary support for this research was provided by Danone Nutricia Research. We further acknowledge support from the Netherlands Organization for Scientific Research (NWO) funding the large-scale proteomics facility Proteins@Work (project 184.032.201) embedded in the Netherlands Proteomics Centre and the TOP-Punt Grant 718.015.003. J.Z. additionally acknowledges support from the Chinese Scholarship Council (CSC) during her PhD at Utrecht University.

Funding Information:
P. van Dam, K. Knipping, M. Mank, and B. Stahl are employees of Danone Nutricia Research. J. Zhu and K.A. Dingess were enrolled as PhD students at Utrecht University during this study and received partial financial support from Danone Nutricia Research. None of the other authors has further conflicts of interest regarding the content of this manuscript.

Publisher Copyright:
© 2021, The Author(s).

Keywords

ELISA
Human milk
Mass spectrometry
Proteomics
sIgA

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Cite this

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title = "Optimization of a human milk-directed quantitative sIgA ELISA method substantiated by mass spectrometry",

abstract = "Immunoglobulins are the primary protective products in human milk and are responsible for transferring maternal pathogen memory to the infant, providing protection by binding to recognized pathogens and inhibiting virulence. To better understand potentially protective/anti-infective compounds in human milk, the establishment of human milk-tailored analytical approaches is crucial, as most contemporary analytical methods have been optimized for plasma or serum. One of the most prominent immunoglobulins in human milk is secretory immunoglobulin A (sIgA), which may be relevant for the protection of breastfed infants from harmful pathogens. Advanced sIgA detection methods can help monitor the immune status and development of the mother-infant dyad. We therefore developed an enzyme-linked immunosorbent assay (ELISA) sIgA method for the quantitative analysis of IgA plus secretory component (SC), validated with sIgA standards and substantiated by mass spectrometry (MS)-based proteomics. A very strong correlation was observed between the MS-detected IgA1 and the human milk-specific sIgA ELISA (r = 0.82). Overall, the MS data indicate that the developed human milk sIgA ELISA does not differentiate between sIgA1 and sIgA2 and is, therefore, a reflection of total sIgA. Furthermore, our MS data and the human milk-derived sIgA ELISA data are better correlated than data derived from a standard serum IgA ELISA kit (relative to MS IgA1 r = 0.82 and r = 0.42, respectively). We therefore propose our human milk-specific sIgA ELISA as an ideal quantitative indicator of total sIgA with advantages over current serum IgA ELISA kits.",

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note = "Funding Information: Primary support for this research was provided by Danone Nutricia Research. We further acknowledge support from the Netherlands Organization for Scientific Research (NWO) funding the large-scale proteomics facility Proteins@Work (project 184.032.201) embedded in the Netherlands Proteomics Centre and the TOP-Punt Grant 718.015.003. J.Z. additionally acknowledges support from the Chinese Scholarship Council (CSC) during her PhD at Utrecht University. Funding Information: P. van Dam, K. Knipping, M. Mank, and B. Stahl are employees of Danone Nutricia Research. J. Zhu and K.A. Dingess were enrolled as PhD students at Utrecht University during this study and received partial financial support from Danone Nutricia Research. None of the other authors has further conflicts of interest regarding the content of this manuscript. Publisher Copyright: {\textcopyright} 2021, The Author(s).",

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AU - van Dam, Pauline

AU - Zhu, Jing

AU - Mank, Marko

AU - Knipping, Karen

AU - Heck, Albert J R

AU - Stahl, Bernd

N1 - Funding Information: Primary support for this research was provided by Danone Nutricia Research. We further acknowledge support from the Netherlands Organization for Scientific Research (NWO) funding the large-scale proteomics facility Proteins@Work (project 184.032.201) embedded in the Netherlands Proteomics Centre and the TOP-Punt Grant 718.015.003. J.Z. additionally acknowledges support from the Chinese Scholarship Council (CSC) during her PhD at Utrecht University. Funding Information: P. van Dam, K. Knipping, M. Mank, and B. Stahl are employees of Danone Nutricia Research. J. Zhu and K.A. Dingess were enrolled as PhD students at Utrecht University during this study and received partial financial support from Danone Nutricia Research. None of the other authors has further conflicts of interest regarding the content of this manuscript. Publisher Copyright: © 2021, The Author(s).

PY - 2021/8

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N2 - Immunoglobulins are the primary protective products in human milk and are responsible for transferring maternal pathogen memory to the infant, providing protection by binding to recognized pathogens and inhibiting virulence. To better understand potentially protective/anti-infective compounds in human milk, the establishment of human milk-tailored analytical approaches is crucial, as most contemporary analytical methods have been optimized for plasma or serum. One of the most prominent immunoglobulins in human milk is secretory immunoglobulin A (sIgA), which may be relevant for the protection of breastfed infants from harmful pathogens. Advanced sIgA detection methods can help monitor the immune status and development of the mother-infant dyad. We therefore developed an enzyme-linked immunosorbent assay (ELISA) sIgA method for the quantitative analysis of IgA plus secretory component (SC), validated with sIgA standards and substantiated by mass spectrometry (MS)-based proteomics. A very strong correlation was observed between the MS-detected IgA1 and the human milk-specific sIgA ELISA (r = 0.82). Overall, the MS data indicate that the developed human milk sIgA ELISA does not differentiate between sIgA1 and sIgA2 and is, therefore, a reflection of total sIgA. Furthermore, our MS data and the human milk-derived sIgA ELISA data are better correlated than data derived from a standard serum IgA ELISA kit (relative to MS IgA1 r = 0.82 and r = 0.42, respectively). We therefore propose our human milk-specific sIgA ELISA as an ideal quantitative indicator of total sIgA with advantages over current serum IgA ELISA kits.

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Optimization of a human milk-directed quantitative sIgA ELISA method substantiated by mass spectrometry

Abstract

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Keywords

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