Pattern variations of polyclonal and monoclonal immunoglobulins of different isotypes analyzed by high-resolution two-dimensional electrophoresis.
High-resolution two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) was used to analyze serum samples and purified immunoglobulins (Ig) obtained from "normal" individuals and from patients diagnosed with monoclonal gammopathies (MG) (n = 47; 5 IgA, 15 IgM, 15 IgG, 4 biclonal IgG, 1 IgD, 7 Bence Jones proteins). Polyclonal and monoclonal heavy (H) chains were located at different restricted gel positions according to their isotype. Monoclonal H chains appeared as sets of spots characterized by charge (pI) and size (M(r)) microheterogeneity. Most of the monoclonal gamma chains were not seen on the gels (12/15). Supplementary polypeptides of 45-48 kDa were detected in serum samples containing monoclonal IgM, but were not seen in MG of other isotypes. However, these polypeptides were not specifically associated with monoclonal IgM because they were also found on protein maps of purified polyclonal IgM. Polyclonal light (L) chains appeared as cloudy bands containing several zones of higher density, whereas monoclonal L chains were usually resolved as single sharp spots. In 6 samples, monoclonal L chains were not seen, and in 9 samples, they appeared as two or more spots, characterized by different pI and/or M(r). In one sample obtained from a patient with a biclonal gammopathy, the L chains were resolved as 4 different spots. Our results confirm that 2-D PAGE is an excellent tool to study Ig. Analysis of the L chain region of the gels was particularly informative. Several monoclonal L chains exhibited heterogeneous two-dimensional spot patterns, suggesting that "subtile" clonal mutations of B-cell lineage and/or posttranslational modifications were involved in their production.
Tissot
J D
JD
Centre de Transfusion Sanguine de la Croix-Rouge suisse, Lausanne, Switzerland.
Hochstrasser
D F
DF
Spertini
F
F
Schifferli
J A
JA
Schneider
P
P
D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D001797:Blood Protein Electrophoresis / Q000379:methods; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D005069:Evaluation Studies as Topic; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000302:isolation & purification; D007132:Immunoglobulin Isotypes / Q000302:isolation & purification*; D007147:Immunoglobulin Light Chains / Q000302:isolation & purification; D007075:Immunoglobulin M / Q000302:isolation & purification; D010265:Paraproteinemias / Q000276:immunology; D010449:Peptide Mapping / Q000379:methods
D000911:Antibodies, Monoclonal; D007143:Immunoglobulin Heavy Chains; D007132:Immunoglobulin Isotypes; D007147:Immunoglobulin Light Chains; D007075:Immunoglobulin M