IgG heavy-chain sublcass typing of myeloma paraproteins by isoelectric focusing immunoblot analysis

F. J. Fasullo, H. A. Fritsche, F. J. Liu, Robert G Hamilton

Research output: Contribution to journalArticle

Abstract

The objective of this study was to develop a clinical laboratory method for subclass typing of human immunoglobulin G (IgG) paraproteins. Serum proteins were isoelectrically focused (IEF) in a minigel and passively blotted by capillary diffusion onto untreated nitrocellulose. Unreacted sites on the nitrocellulose were blocked with bovine serum albumin and the bound IgG was detected with peroxidase-conjugated anti-human IgG1-4 monoclonal antibodies from WHO/IUIS clones. The IEF immunoblot specificity was demonstrated by analysis of documented IgG, IgA, and IgM myeloma proteins of known subclass and light-chain composition. IEF immunoblots of sera from 18 myeloma patients who had an above-normal total IgG concentration produced IEF immunoblot patterns composed of five to 10 discrete bands (pI range 6.0 to 8.4). In contrast, no detectable IgG bands were observed with sera containing IgA and IgM paraproteins. The observed subclass frequencies of IgG paraproteins were 56% IgG1 (10/18), 28% IgG2 (5/18), 11% IgG3 (2/18), and 5% IgG4 (1/18). IEF immunoblot analysis permits the monitoring of changes in the pI and subclass of an IgG paraprotein over the course of a myeloma patient's therapy program.

Original languageEnglish (US)
Pages (from-to)364-368
Number of pages5
JournalClinical Chemistry
Volume35
Issue number3
StatePublished - 1989
Externally publishedYes

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Paraproteins
Immunoglobulin Heavy Chains
Isoelectric Focusing
Immunoglobulin G
Collodion
Immunoglobulin A
Immunoglobulin M
Clinical laboratories
Myeloma Proteins
Bovine Serum Albumin
Serum
Peroxidase
Blood Proteins

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this

IgG heavy-chain sublcass typing of myeloma paraproteins by isoelectric focusing immunoblot analysis. / Fasullo, F. J.; Fritsche, H. A.; Liu, F. J.; Hamilton, Robert G.

In: Clinical Chemistry, Vol. 35, No. 3, 1989, p. 364-368.

Research output: Contribution to journalArticle

Fasullo, F. J. ; Fritsche, H. A. ; Liu, F. J. ; Hamilton, Robert G. / IgG heavy-chain sublcass typing of myeloma paraproteins by isoelectric focusing immunoblot analysis. In: Clinical Chemistry. 1989 ; Vol. 35, No. 3. pp. 364-368.
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