Characterization of amyloidogenic immunoglobulin light chains directly from serum by on-line immunoaffinity isolation

Harold Robert (Bob) III Bergen, Roshini S. Abraham, Kenneth L. Johnson, Arthur R. Bradwell, Stephen Naylor

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Primary systemic amyloidosis (AL) is characterized by the overproduction of immunoglobulin light chain proteins by a monoclonal, terminally differentiated B-lymphocyte or plasma cell clone. The free immunoglobulin light chains are deposited in an abnormal conformation as amyloid in a variety of organs in the body. The mechanism of amyloid formation is not well understood, but appears to be associated with some form of cleavage of the immunoglobulin light chain with subsequent aggregate formation. In an attempt to characterize the structure of amyloid-forming light chain proteins we developed an on-line immunoaffinity purification and subsequent characterization of free κ and free λ immunoglobulin light chains by electrospray ionization mass spectrometry. The methodology is totally automated and requires 20 μL of serum. Mass spectral analysis of Bence Jones proteins under non-denaturing conditions was also utilized to examine the tertiary and quaternary structure of light chain proteins and clearly shows covalent dimer formation of lambda type light chain. This type of on-line assay may prove helpful in elucidating distinguishing features capable of discriminating AL from benign monoclonal gammopathies of undetermined significance as well as diagnosing AL.

Original languageEnglish (US)
Pages (from-to)191-201
Number of pages11
JournalBiomedical Chromatography
Volume18
Issue number3
DOIs
StatePublished - Apr 2004

Fingerprint

Immunoglobulin Light Chains
Amyloid
Monoclonal Gammopathy of Undetermined Significance
Amyloidosis
Serum
Light
Clone cells
Bence Jones Protein
Electrospray ionization
Proteins
Lymphocytes
Electrospray Ionization Mass Spectrometry
Plasma Cells
Dimers
Spectrum analysis
Purification
Mass spectrometry
Conformations
Assays
B-Lymphocytes

Keywords

  • Desalting trap
  • ESI MS
  • Immunoaffinity cartridge

ASJC Scopus subject areas

  • Analytical Chemistry
  • Clinical Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Pharmacology

Cite this

Characterization of amyloidogenic immunoglobulin light chains directly from serum by on-line immunoaffinity isolation. / Bergen, Harold Robert (Bob) III; Abraham, Roshini S.; Johnson, Kenneth L.; Bradwell, Arthur R.; Naylor, Stephen.

In: Biomedical Chromatography, Vol. 18, No. 3, 04.2004, p. 191-201.

Research output: Contribution to journalArticle

Bergen, Harold Robert (Bob) III ; Abraham, Roshini S. ; Johnson, Kenneth L. ; Bradwell, Arthur R. ; Naylor, Stephen. / Characterization of amyloidogenic immunoglobulin light chains directly from serum by on-line immunoaffinity isolation. In: Biomedical Chromatography. 2004 ; Vol. 18, No. 3. pp. 191-201.
@article{e66b625730864a7cb48b4630d5416f19,
title = "Characterization of amyloidogenic immunoglobulin light chains directly from serum by on-line immunoaffinity isolation",
abstract = "Primary systemic amyloidosis (AL) is characterized by the overproduction of immunoglobulin light chain proteins by a monoclonal, terminally differentiated B-lymphocyte or plasma cell clone. The free immunoglobulin light chains are deposited in an abnormal conformation as amyloid in a variety of organs in the body. The mechanism of amyloid formation is not well understood, but appears to be associated with some form of cleavage of the immunoglobulin light chain with subsequent aggregate formation. In an attempt to characterize the structure of amyloid-forming light chain proteins we developed an on-line immunoaffinity purification and subsequent characterization of free κ and free λ immunoglobulin light chains by electrospray ionization mass spectrometry. The methodology is totally automated and requires 20 μL of serum. Mass spectral analysis of Bence Jones proteins under non-denaturing conditions was also utilized to examine the tertiary and quaternary structure of light chain proteins and clearly shows covalent dimer formation of lambda type light chain. This type of on-line assay may prove helpful in elucidating distinguishing features capable of discriminating AL from benign monoclonal gammopathies of undetermined significance as well as diagnosing AL.",
keywords = "Desalting trap, ESI MS, Immunoaffinity cartridge",
author = "Bergen, {Harold Robert (Bob) III} and Abraham, {Roshini S.} and Johnson, {Kenneth L.} and Bradwell, {Arthur R.} and Stephen Naylor",
year = "2004",
month = "4",
doi = "10.1002/bmc.323",
language = "English (US)",
volume = "18",
pages = "191--201",
journal = "Biomedical Chromatography",
issn = "0269-3879",
publisher = "John Wiley and Sons Ltd",
number = "3",

}

TY - JOUR

T1 - Characterization of amyloidogenic immunoglobulin light chains directly from serum by on-line immunoaffinity isolation

AU - Bergen, Harold Robert (Bob) III

AU - Abraham, Roshini S.

AU - Johnson, Kenneth L.

AU - Bradwell, Arthur R.

AU - Naylor, Stephen

PY - 2004/4

Y1 - 2004/4

N2 - Primary systemic amyloidosis (AL) is characterized by the overproduction of immunoglobulin light chain proteins by a monoclonal, terminally differentiated B-lymphocyte or plasma cell clone. The free immunoglobulin light chains are deposited in an abnormal conformation as amyloid in a variety of organs in the body. The mechanism of amyloid formation is not well understood, but appears to be associated with some form of cleavage of the immunoglobulin light chain with subsequent aggregate formation. In an attempt to characterize the structure of amyloid-forming light chain proteins we developed an on-line immunoaffinity purification and subsequent characterization of free κ and free λ immunoglobulin light chains by electrospray ionization mass spectrometry. The methodology is totally automated and requires 20 μL of serum. Mass spectral analysis of Bence Jones proteins under non-denaturing conditions was also utilized to examine the tertiary and quaternary structure of light chain proteins and clearly shows covalent dimer formation of lambda type light chain. This type of on-line assay may prove helpful in elucidating distinguishing features capable of discriminating AL from benign monoclonal gammopathies of undetermined significance as well as diagnosing AL.

AB - Primary systemic amyloidosis (AL) is characterized by the overproduction of immunoglobulin light chain proteins by a monoclonal, terminally differentiated B-lymphocyte or plasma cell clone. The free immunoglobulin light chains are deposited in an abnormal conformation as amyloid in a variety of organs in the body. The mechanism of amyloid formation is not well understood, but appears to be associated with some form of cleavage of the immunoglobulin light chain with subsequent aggregate formation. In an attempt to characterize the structure of amyloid-forming light chain proteins we developed an on-line immunoaffinity purification and subsequent characterization of free κ and free λ immunoglobulin light chains by electrospray ionization mass spectrometry. The methodology is totally automated and requires 20 μL of serum. Mass spectral analysis of Bence Jones proteins under non-denaturing conditions was also utilized to examine the tertiary and quaternary structure of light chain proteins and clearly shows covalent dimer formation of lambda type light chain. This type of on-line assay may prove helpful in elucidating distinguishing features capable of discriminating AL from benign monoclonal gammopathies of undetermined significance as well as diagnosing AL.

KW - Desalting trap

KW - ESI MS

KW - Immunoaffinity cartridge

UR - http://www.scopus.com/inward/record.url?scp=2342653503&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=2342653503&partnerID=8YFLogxK

U2 - 10.1002/bmc.323

DO - 10.1002/bmc.323

M3 - Article

C2 - 15103706

AN - SCOPUS:2342653503

VL - 18

SP - 191

EP - 201

JO - Biomedical Chromatography

JF - Biomedical Chromatography

SN - 0269-3879

IS - 3

ER -