Phenotyping polyclonal kappa and lambda light chain molecular mass distributions in patient serum using mass spectrometry

David R. Barnidge, Surendra Dasari, Marina Ramirez-Alvarado, Adrian Fontan, Maria A V Willrich, Renee C. Tschumper, Diane F Jelinek, Melissa R. Snyder, Angela Dispenzieri, Jerry A. Katzmann, David L. Murray

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

We previously described a microLC-ESI-Q-TOF MS method for identifying monoclonal immunoglobulins in serum and then tracking them over time using their accurate molecular mass. Here we demonstrate how the same methodology can be used to identify and characterize polyclonal immunoglobulins in serum. We establish that two molecular mass distributions observed by microLC-ESI-Q-TOF MS are from polyclonal kappa and lambda light chains using a combination of theoretical molecular masses from gene sequence data and the analysis of commercially available purified polyclonal IgG kappa and IgG lambda from normal human serum. A linear regression comparison of kappa/lambda ratios for 74 serum samples (25 hypergammaglobulinemia, 24 hypogammaglobulinemia, 25 normal) determined by microflowLC-ESI-Q-TOF MS and immunonephelometry had a slope of 1.37 and a correlation coefficient of 0.639. In addition to providing kappa/lambda ratios, the same microLC-ESI-Q-TOF MS analysis can determine the molecular mass for oligoclonal light chains observed above the polyclonal background in patient samples. In 2 patients with immune disorders and hypergammaglobulinemia, we observed a skewed polyclonal molecular mass distribution which translated into biased kappa/lambda ratios. Mass spectrometry provides a rapid and simple way to combine the polyclonal kappa/lambda light chain abundance ratios with the identification of dominant monoclonal as well as oligoclonal light chain immunoglobulins. We anticipate that this approach to evaluating immunoglobulin light chains will lead to improved understanding of immune deficiencies, autoimmune diseases, and antibody responses.

Original languageEnglish (US)
Pages (from-to)5198-5205
Number of pages8
JournalJournal of Proteome Research
Volume13
Issue number11
DOIs
StatePublished - Nov 7 2014

Fingerprint

Molecular mass
Mass spectrometry
Mass Spectrometry
Hypergammaglobulinemia
Immunoglobulin Light Chains
Light
Serum
Immunoglobulins
Immunoglobulin G
Oligoclonal Bands
Agammaglobulinemia
Immune System Diseases
Autoimmunity
Autoimmune Diseases
Antibody Formation
Sequence Analysis
Linear Models
Linear regression
Genes
Antibodies

Keywords

  • autoimmune diseases
  • Immunoglobulins
  • kappa/lambda ratios
  • monoclonal
  • polyclonal

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

Cite this

Phenotyping polyclonal kappa and lambda light chain molecular mass distributions in patient serum using mass spectrometry. / Barnidge, David R.; Dasari, Surendra; Ramirez-Alvarado, Marina; Fontan, Adrian; Willrich, Maria A V; Tschumper, Renee C.; Jelinek, Diane F; Snyder, Melissa R.; Dispenzieri, Angela; Katzmann, Jerry A.; Murray, David L.

In: Journal of Proteome Research, Vol. 13, No. 11, 07.11.2014, p. 5198-5205.

Research output: Contribution to journalArticle

Barnidge, David R. ; Dasari, Surendra ; Ramirez-Alvarado, Marina ; Fontan, Adrian ; Willrich, Maria A V ; Tschumper, Renee C. ; Jelinek, Diane F ; Snyder, Melissa R. ; Dispenzieri, Angela ; Katzmann, Jerry A. ; Murray, David L. / Phenotyping polyclonal kappa and lambda light chain molecular mass distributions in patient serum using mass spectrometry. In: Journal of Proteome Research. 2014 ; Vol. 13, No. 11. pp. 5198-5205.
@article{7bad4ec6eb044ad8b03f7682da74e857,
title = "Phenotyping polyclonal kappa and lambda light chain molecular mass distributions in patient serum using mass spectrometry",
abstract = "We previously described a microLC-ESI-Q-TOF MS method for identifying monoclonal immunoglobulins in serum and then tracking them over time using their accurate molecular mass. Here we demonstrate how the same methodology can be used to identify and characterize polyclonal immunoglobulins in serum. We establish that two molecular mass distributions observed by microLC-ESI-Q-TOF MS are from polyclonal kappa and lambda light chains using a combination of theoretical molecular masses from gene sequence data and the analysis of commercially available purified polyclonal IgG kappa and IgG lambda from normal human serum. A linear regression comparison of kappa/lambda ratios for 74 serum samples (25 hypergammaglobulinemia, 24 hypogammaglobulinemia, 25 normal) determined by microflowLC-ESI-Q-TOF MS and immunonephelometry had a slope of 1.37 and a correlation coefficient of 0.639. In addition to providing kappa/lambda ratios, the same microLC-ESI-Q-TOF MS analysis can determine the molecular mass for oligoclonal light chains observed above the polyclonal background in patient samples. In 2 patients with immune disorders and hypergammaglobulinemia, we observed a skewed polyclonal molecular mass distribution which translated into biased kappa/lambda ratios. Mass spectrometry provides a rapid and simple way to combine the polyclonal kappa/lambda light chain abundance ratios with the identification of dominant monoclonal as well as oligoclonal light chain immunoglobulins. We anticipate that this approach to evaluating immunoglobulin light chains will lead to improved understanding of immune deficiencies, autoimmune diseases, and antibody responses.",
keywords = "autoimmune diseases, Immunoglobulins, kappa/lambda ratios, monoclonal, polyclonal",
author = "Barnidge, {David R.} and Surendra Dasari and Marina Ramirez-Alvarado and Adrian Fontan and Willrich, {Maria A V} and Tschumper, {Renee C.} and Jelinek, {Diane F} and Snyder, {Melissa R.} and Angela Dispenzieri and Katzmann, {Jerry A.} and Murray, {David L.}",
year = "2014",
month = "11",
day = "7",
doi = "10.1021/pr5005967",
language = "English (US)",
volume = "13",
pages = "5198--5205",
journal = "Journal of Proteome Research",
issn = "1535-3893",
publisher = "American Chemical Society",
number = "11",

}

TY - JOUR

T1 - Phenotyping polyclonal kappa and lambda light chain molecular mass distributions in patient serum using mass spectrometry

AU - Barnidge, David R.

AU - Dasari, Surendra

AU - Ramirez-Alvarado, Marina

AU - Fontan, Adrian

AU - Willrich, Maria A V

AU - Tschumper, Renee C.

AU - Jelinek, Diane F

AU - Snyder, Melissa R.

AU - Dispenzieri, Angela

AU - Katzmann, Jerry A.

AU - Murray, David L.

PY - 2014/11/7

Y1 - 2014/11/7

N2 - We previously described a microLC-ESI-Q-TOF MS method for identifying monoclonal immunoglobulins in serum and then tracking them over time using their accurate molecular mass. Here we demonstrate how the same methodology can be used to identify and characterize polyclonal immunoglobulins in serum. We establish that two molecular mass distributions observed by microLC-ESI-Q-TOF MS are from polyclonal kappa and lambda light chains using a combination of theoretical molecular masses from gene sequence data and the analysis of commercially available purified polyclonal IgG kappa and IgG lambda from normal human serum. A linear regression comparison of kappa/lambda ratios for 74 serum samples (25 hypergammaglobulinemia, 24 hypogammaglobulinemia, 25 normal) determined by microflowLC-ESI-Q-TOF MS and immunonephelometry had a slope of 1.37 and a correlation coefficient of 0.639. In addition to providing kappa/lambda ratios, the same microLC-ESI-Q-TOF MS analysis can determine the molecular mass for oligoclonal light chains observed above the polyclonal background in patient samples. In 2 patients with immune disorders and hypergammaglobulinemia, we observed a skewed polyclonal molecular mass distribution which translated into biased kappa/lambda ratios. Mass spectrometry provides a rapid and simple way to combine the polyclonal kappa/lambda light chain abundance ratios with the identification of dominant monoclonal as well as oligoclonal light chain immunoglobulins. We anticipate that this approach to evaluating immunoglobulin light chains will lead to improved understanding of immune deficiencies, autoimmune diseases, and antibody responses.

AB - We previously described a microLC-ESI-Q-TOF MS method for identifying monoclonal immunoglobulins in serum and then tracking them over time using their accurate molecular mass. Here we demonstrate how the same methodology can be used to identify and characterize polyclonal immunoglobulins in serum. We establish that two molecular mass distributions observed by microLC-ESI-Q-TOF MS are from polyclonal kappa and lambda light chains using a combination of theoretical molecular masses from gene sequence data and the analysis of commercially available purified polyclonal IgG kappa and IgG lambda from normal human serum. A linear regression comparison of kappa/lambda ratios for 74 serum samples (25 hypergammaglobulinemia, 24 hypogammaglobulinemia, 25 normal) determined by microflowLC-ESI-Q-TOF MS and immunonephelometry had a slope of 1.37 and a correlation coefficient of 0.639. In addition to providing kappa/lambda ratios, the same microLC-ESI-Q-TOF MS analysis can determine the molecular mass for oligoclonal light chains observed above the polyclonal background in patient samples. In 2 patients with immune disorders and hypergammaglobulinemia, we observed a skewed polyclonal molecular mass distribution which translated into biased kappa/lambda ratios. Mass spectrometry provides a rapid and simple way to combine the polyclonal kappa/lambda light chain abundance ratios with the identification of dominant monoclonal as well as oligoclonal light chain immunoglobulins. We anticipate that this approach to evaluating immunoglobulin light chains will lead to improved understanding of immune deficiencies, autoimmune diseases, and antibody responses.

KW - autoimmune diseases

KW - Immunoglobulins

KW - kappa/lambda ratios

KW - monoclonal

KW - polyclonal

UR - http://www.scopus.com/inward/record.url?scp=84908894078&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84908894078&partnerID=8YFLogxK

U2 - 10.1021/pr5005967

DO - 10.1021/pr5005967

M3 - Article

C2 - 25134970

AN - SCOPUS:84908894078

VL - 13

SP - 5198

EP - 5205

JO - Journal of Proteome Research

JF - Journal of Proteome Research

SN - 1535-3893

IS - 11

ER -