Comprehensive assessment of M-proteins using nanobody enrichment coupled to MALDI-TOF mass spectrometry

John R. Mills, Mindy C. Kohlhagen, Surendra Dasari, Patrick M. Vanderboom, Robert A. Kyle, Jerry A. Katzmann, Maria A V Willrich, David R. Barnidge, Angela Dispenzieri, David L. Murray

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

BACKGROUND: Electrophoretic separation of serum and urine proteins has played a central role in diagnosing and monitoring plasma cell disorders. Despite limitations in resolution and analytical sensitivity, plus the necessity for adjunct methods, protein gel electrophoresis and immunofixation electrophoresis (IFE) remain front-line tests. METHODS: We developed a MALDI mass spectrometry-based assay that was simple to perform, automatable, analytically sensitive, and applicable to analyzing the wide variety of monoclonal proteins (M-proteins) encountered clinically. This assay, called MASS-FIX, used the unique molecular mass signatures of the different Ig isotypes in combination with nanobody immunoenrichment to generate information-rich mass spectra from which M-proteins could be identified, isotyped, and quantified. The performance of MASS-FIX was compared to current gel-based electrophoresis assays. RESULTS: MASS-FIX detected all M-proteins that were detectable by urine or serum protein electrophoresis. In serial dilution studies, MASS-FIX was more analytically sensitive than IFE. For patient samples, MASS-FIX provided the same primary isotype information for 98% of serum M-proteins (n = 152) and 95% of urine M-proteins (n = 55). MASS-FIX accurately quantified M-protein to <1 g/dL, with reduced bias as compared to protein electrophoresis. Intraassay and interassay CVs were <20% across all samples having M-protein concentrations >0.045 g/dL, with the ability to detect M-proteins <0.01 g/dL. In addition, MASS-FIX could simultaneously measure κ:λ light chain ratios for IgG, IgA, and IgM. Retrospective serial monitoring of patients with myeloma posttreatment demonstrated that MASS-FIX provided equivalent quantitative information to either protein electrophoresis or the Hevylite™ assay. CONCLUSIONS: MASS-FIX can advance how plasma cell disorders are screened, diagnosed, and monitored.

Original languageEnglish (US)
Pages (from-to)1334-1344
Number of pages11
JournalClinical Chemistry
Volume62
Issue number10
DOIs
StatePublished - Oct 1 2016

Fingerprint

Single-Domain Antibodies
Matrix-Assisted Laser Desorption-Ionization Mass Spectrometry
Mass spectrometry
Mass Spectrometry
Electrophoresis
Proteins
Assays
Blood Proteins
Urine
Plasma Cells
Gels
Physiologic Monitoring
Plasmas
Immunoglobulin A
Monitoring
Immunoglobulin M
Molecular mass
Immunoglobulin G
Dilution
Light

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, medical

Cite this

Mills, J. R., Kohlhagen, M. C., Dasari, S., Vanderboom, P. M., Kyle, R. A., Katzmann, J. A., ... Murray, D. L. (2016). Comprehensive assessment of M-proteins using nanobody enrichment coupled to MALDI-TOF mass spectrometry. Clinical Chemistry, 62(10), 1334-1344. https://doi.org/10.1373/clinchem.2015.253740

Comprehensive assessment of M-proteins using nanobody enrichment coupled to MALDI-TOF mass spectrometry. / Mills, John R.; Kohlhagen, Mindy C.; Dasari, Surendra; Vanderboom, Patrick M.; Kyle, Robert A.; Katzmann, Jerry A.; Willrich, Maria A V; Barnidge, David R.; Dispenzieri, Angela; Murray, David L.

In: Clinical Chemistry, Vol. 62, No. 10, 01.10.2016, p. 1334-1344.

Research output: Contribution to journalArticle

Mills, JR, Kohlhagen, MC, Dasari, S, Vanderboom, PM, Kyle, RA, Katzmann, JA, Willrich, MAV, Barnidge, DR, Dispenzieri, A & Murray, DL 2016, 'Comprehensive assessment of M-proteins using nanobody enrichment coupled to MALDI-TOF mass spectrometry', Clinical Chemistry, vol. 62, no. 10, pp. 1334-1344. https://doi.org/10.1373/clinchem.2015.253740
Mills, John R. ; Kohlhagen, Mindy C. ; Dasari, Surendra ; Vanderboom, Patrick M. ; Kyle, Robert A. ; Katzmann, Jerry A. ; Willrich, Maria A V ; Barnidge, David R. ; Dispenzieri, Angela ; Murray, David L. / Comprehensive assessment of M-proteins using nanobody enrichment coupled to MALDI-TOF mass spectrometry. In: Clinical Chemistry. 2016 ; Vol. 62, No. 10. pp. 1334-1344.
@article{7ca4f6545ee347cc89b7f9ea221ca537,
title = "Comprehensive assessment of M-proteins using nanobody enrichment coupled to MALDI-TOF mass spectrometry",
abstract = "BACKGROUND: Electrophoretic separation of serum and urine proteins has played a central role in diagnosing and monitoring plasma cell disorders. Despite limitations in resolution and analytical sensitivity, plus the necessity for adjunct methods, protein gel electrophoresis and immunofixation electrophoresis (IFE) remain front-line tests. METHODS: We developed a MALDI mass spectrometry-based assay that was simple to perform, automatable, analytically sensitive, and applicable to analyzing the wide variety of monoclonal proteins (M-proteins) encountered clinically. This assay, called MASS-FIX, used the unique molecular mass signatures of the different Ig isotypes in combination with nanobody immunoenrichment to generate information-rich mass spectra from which M-proteins could be identified, isotyped, and quantified. The performance of MASS-FIX was compared to current gel-based electrophoresis assays. RESULTS: MASS-FIX detected all M-proteins that were detectable by urine or serum protein electrophoresis. In serial dilution studies, MASS-FIX was more analytically sensitive than IFE. For patient samples, MASS-FIX provided the same primary isotype information for 98{\%} of serum M-proteins (n = 152) and 95{\%} of urine M-proteins (n = 55). MASS-FIX accurately quantified M-protein to <1 g/dL, with reduced bias as compared to protein electrophoresis. Intraassay and interassay CVs were <20{\%} across all samples having M-protein concentrations >0.045 g/dL, with the ability to detect M-proteins <0.01 g/dL. In addition, MASS-FIX could simultaneously measure κ:λ light chain ratios for IgG, IgA, and IgM. Retrospective serial monitoring of patients with myeloma posttreatment demonstrated that MASS-FIX provided equivalent quantitative information to either protein electrophoresis or the Hevylite™ assay. CONCLUSIONS: MASS-FIX can advance how plasma cell disorders are screened, diagnosed, and monitored.",
author = "Mills, {John R.} and Kohlhagen, {Mindy C.} and Surendra Dasari and Vanderboom, {Patrick M.} and Kyle, {Robert A.} and Katzmann, {Jerry A.} and Willrich, {Maria A V} and Barnidge, {David R.} and Angela Dispenzieri and Murray, {David L.}",
year = "2016",
month = "10",
day = "1",
doi = "10.1373/clinchem.2015.253740",
language = "English (US)",
volume = "62",
pages = "1334--1344",
journal = "Clinical Chemistry",
issn = "0009-9147",
publisher = "American Association for Clinical Chemistry Inc.",
number = "10",

}

TY - JOUR

T1 - Comprehensive assessment of M-proteins using nanobody enrichment coupled to MALDI-TOF mass spectrometry

AU - Mills, John R.

AU - Kohlhagen, Mindy C.

AU - Dasari, Surendra

AU - Vanderboom, Patrick M.

AU - Kyle, Robert A.

AU - Katzmann, Jerry A.

AU - Willrich, Maria A V

AU - Barnidge, David R.

AU - Dispenzieri, Angela

AU - Murray, David L.

PY - 2016/10/1

Y1 - 2016/10/1

N2 - BACKGROUND: Electrophoretic separation of serum and urine proteins has played a central role in diagnosing and monitoring plasma cell disorders. Despite limitations in resolution and analytical sensitivity, plus the necessity for adjunct methods, protein gel electrophoresis and immunofixation electrophoresis (IFE) remain front-line tests. METHODS: We developed a MALDI mass spectrometry-based assay that was simple to perform, automatable, analytically sensitive, and applicable to analyzing the wide variety of monoclonal proteins (M-proteins) encountered clinically. This assay, called MASS-FIX, used the unique molecular mass signatures of the different Ig isotypes in combination with nanobody immunoenrichment to generate information-rich mass spectra from which M-proteins could be identified, isotyped, and quantified. The performance of MASS-FIX was compared to current gel-based electrophoresis assays. RESULTS: MASS-FIX detected all M-proteins that were detectable by urine or serum protein electrophoresis. In serial dilution studies, MASS-FIX was more analytically sensitive than IFE. For patient samples, MASS-FIX provided the same primary isotype information for 98% of serum M-proteins (n = 152) and 95% of urine M-proteins (n = 55). MASS-FIX accurately quantified M-protein to <1 g/dL, with reduced bias as compared to protein electrophoresis. Intraassay and interassay CVs were <20% across all samples having M-protein concentrations >0.045 g/dL, with the ability to detect M-proteins <0.01 g/dL. In addition, MASS-FIX could simultaneously measure κ:λ light chain ratios for IgG, IgA, and IgM. Retrospective serial monitoring of patients with myeloma posttreatment demonstrated that MASS-FIX provided equivalent quantitative information to either protein electrophoresis or the Hevylite™ assay. CONCLUSIONS: MASS-FIX can advance how plasma cell disorders are screened, diagnosed, and monitored.

AB - BACKGROUND: Electrophoretic separation of serum and urine proteins has played a central role in diagnosing and monitoring plasma cell disorders. Despite limitations in resolution and analytical sensitivity, plus the necessity for adjunct methods, protein gel electrophoresis and immunofixation electrophoresis (IFE) remain front-line tests. METHODS: We developed a MALDI mass spectrometry-based assay that was simple to perform, automatable, analytically sensitive, and applicable to analyzing the wide variety of monoclonal proteins (M-proteins) encountered clinically. This assay, called MASS-FIX, used the unique molecular mass signatures of the different Ig isotypes in combination with nanobody immunoenrichment to generate information-rich mass spectra from which M-proteins could be identified, isotyped, and quantified. The performance of MASS-FIX was compared to current gel-based electrophoresis assays. RESULTS: MASS-FIX detected all M-proteins that were detectable by urine or serum protein electrophoresis. In serial dilution studies, MASS-FIX was more analytically sensitive than IFE. For patient samples, MASS-FIX provided the same primary isotype information for 98% of serum M-proteins (n = 152) and 95% of urine M-proteins (n = 55). MASS-FIX accurately quantified M-protein to <1 g/dL, with reduced bias as compared to protein electrophoresis. Intraassay and interassay CVs were <20% across all samples having M-protein concentrations >0.045 g/dL, with the ability to detect M-proteins <0.01 g/dL. In addition, MASS-FIX could simultaneously measure κ:λ light chain ratios for IgG, IgA, and IgM. Retrospective serial monitoring of patients with myeloma posttreatment demonstrated that MASS-FIX provided equivalent quantitative information to either protein electrophoresis or the Hevylite™ assay. CONCLUSIONS: MASS-FIX can advance how plasma cell disorders are screened, diagnosed, and monitored.

UR - http://www.scopus.com/inward/record.url?scp=84990047383&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84990047383&partnerID=8YFLogxK

U2 - 10.1373/clinchem.2015.253740

DO - 10.1373/clinchem.2015.253740

M3 - Article

VL - 62

SP - 1334

EP - 1344

JO - Clinical Chemistry

JF - Clinical Chemistry

SN - 0009-9147

IS - 10

ER -