Two-dimensional polyacrylamide gel electrophoresis analysis of cryoglobulins and identification of an IgM-associated peptide

Details

Serval ID
serval:BIB_C9574665858C
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
Two-dimensional polyacrylamide gel electrophoresis analysis of cryoglobulins and identification of an IgM-associated peptide
Journal
Journal of Immunological Methods
Author(s)
Tissot  J. D., Schifferli  J. A., Hochstrasser  D. F., Pasquali  C., Spertini  F., Clement  F., Frutiger  S., Paquet  N., Hughes  G. J., Schneider  P.
ISSN
0022-1759 (Print)
Publication state
Published
Issued date
07/1994
Volume
173
Number
1
Pages
63-75
Notes
Journal Article
Research Support, Non-U.S. Gov't --- Old month value: Jul 12
Abstract
The clonality of immunoglobulins (Igs) in cryoprecipitates (n = 41) was studied by two-dimensional polyacrylamide gel electrophoresis (2-D PAGE). Our series included 24 cryoglobulins characterized by immunofixation electrophoresis (IF), 12 'trace amount' cryoglobulins, defined by a protein content in the precipitate of less than 0.05 mg/ml of serum, and five cryoglobulins of undetermined protein composition by IF. 2-D PAGE analysis showed polyclonal IgG associated either with monoclonal Igs (type II cryoglobulins; n = 14) or with polyclonal IgM (type III cryoglobulins; n = 14). In ten cryoprecipitates (two 'trace amount' cryoglobulins as well as seven of 19 type II and as one of five type III cryoglobulins by IF) polyclonal IgG were associated with a mixture of polyclonal and monoclonal IgM. These cryoglobulins were tentatively named type II-III cryoglobulins. A monoclonal IgM was observed in one cryoprecipitate (type I cryoglobulins). Two cryoglobulins presented unexpected 2-D patterns, characterized by the presence of oligoclonal IgM, with trace amounts of Igs of different isotypes (tentatively named type II-III(variant) cryoglobulins). A peptide of 44 kDa with a pI of 5.45 was observed in all cryoglobulins containing IgM (n = 40). This peptide was also present in purified monoclonal or polyclonal IgM fractions. N-terminal microsequencing (12 amino acid residues) revealed that this IgM-associated peptide was an unknown protein. Our results highlight the role of 2-D PAGE as an aid in the analysis of cryoglobulins.
Keywords
Amino Acid Sequence Antibodies, Monoclonal/blood Cryoglobulinemia/blood/immunology Cryoglobulins/*analysis/classification Electrophoresis, Gel, Two-Dimensional/*methods Humans Immunoglobulin Isotypes/blood Immunoglobulin M/*blood Isoelectric Point Molecular Sequence Data Molecular Weight Peptides/*blood/chemistry/genetics
Pubmed
Web of science
Create date
25/01/2008 15:34
Last modification date
20/08/2019 15:44
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