
Full text loading...
Category: Immunology
Immunochemical Characterization of Immunoglobulins in Serum, Urine, and Cerebrospinal Fluid, Page 1 of 2
< Previous page | Next page > /docserver/preview/fulltext/10.1128/9781555818722/9781555818715_CH09-1.gif /docserver/preview/fulltext/10.1128/9781555818722/9781555818715_CH09-2.gifAbstract:
The characterization of immunoglobulins spans a spectrum of methods, including molecular analysis of gene usage and rearrangement, quantitation of immunoglobulin heavy chains as well as intact and free light chains, qualitative assessment and characterization of clonality, and identification of abnormalities that may be clinically significant, such as hyperviscosity syndrome, cryoglobulinemia, and amyloidosis (AL). This chapter focuses on qualitative methods for the assessment and characterization of clonality. The methods include agarose gel electrophoresis (AGE) with immunofixation, capillary electrophoresis (CE) with immunosubtraction (ISUB), and isoelectric focusing with immunoblotting or immunofixation. All three methods can be used to identify monoclonal, oligoclonal, and polyclonal immunoglobulin populations and to identify the heavy and/or light chains contained in the population. Immunofixation electrophoresis (IFE) and ISUB electrophoresis are diagnostic tools used for the identification of monoclonal gammopathies and, conversely, for the confirmation of polyclonal hypergammaglobulinemia. Isoelectric focusing with immunoblotting or immunofixation is a cerebrospinal fluid (CSF) diagnostic test for the identification of oligoclonal bands in multiple sclerosis (MS).
Full text loading...
(A) Normal serum electropherogram; (B) normal serum immunofixation gel.
(A) Normal serum electropherogram; (B) normal serum immunofixation gel.
(A) Electropherogram with monoclonal protein in the gamma region; (B) immunofixation gel showing an IgG lambda monoclonal protein.
(A) Electropherogram with monoclonal protein in the gamma region; (B) immunofixation gel showing an IgG lambda monoclonal protein.
Serum immunofixation gel with no staining in the lambda lane due to lambda antiserum not being applied.
Serum immunofixation gel with no staining in the lambda lane due to lambda antiserum not being applied.
Normal serum immunosubtraction pattern.
Normal serum immunosubtraction pattern.
Serum immunosubtraction pattern with IgG lambda monoclonal protein.
Serum immunosubtraction pattern with IgG lambda monoclonal protein.
(A) Electropherogram with polyclonal increase in gamma globulins; (B) immunofixation gel with polyclonal increase in IgG (a “fuzzy” band is noted in the lambda lane).
(A) Electropherogram with polyclonal increase in gamma globulins; (B) immunofixation gel with polyclonal increase in IgG (a “fuzzy” band is noted in the lambda lane).
Polyclonal increase in an IgG4 subclass. (A) Electropherogram with a rounded peak in the beta-gamma region; (B) immunofixation gel showing an increase in staining in the beta-gamma region in the IgG and both kappa and lambda lanes.
Polyclonal increase in an IgG4 subclass. (A) Electropherogram with a rounded peak in the beta-gamma region; (B) immunofixation gel showing an increase in staining in the beta-gamma region in the IgG and both kappa and lambda lanes.
Serum immunofixation gel with a biclonal gammopathy (IgG lambda and IgA kappa). Antigen excess is also present in the IgA and kappa lanes.
Serum immunofixation gel with a biclonal gammopathy (IgG lambda and IgA kappa). Antigen excess is also present in the IgA and kappa lanes.
Serum immunofixation gel with a free lambda monoclonal protein.
Serum immunofixation gel with a free lambda monoclonal protein.
IgG heavy-chain disease. (A) Immunosubtraction showing removal of only IgG from the small gamma peak (no change is seen with kappa and lambda antisera); (B) immunofixation gel with only an IgG band.
IgG heavy-chain disease. (A) Immunosubtraction showing removal of only IgG from the small gamma peak (no change is seen with kappa and lambda antisera); (B) immunofixation gel with only an IgG band.
Prominent oligoclonal banding. (A) Immunosubtraction; (B) immunofixation gel.
Prominent oligoclonal banding. (A) Immunosubtraction; (B) immunofixation gel.
Contrast media artifact. (A) Electropherogram from capillary electrophoresis showing a peak (arrow) in the alpha-2/beta region; (B) immunofixation gel with no band corresponding to the contrast medium peak.
Contrast media artifact. (A) Electropherogram from capillary electrophoresis showing a peak (arrow) in the alpha-2/beta region; (B) immunofixation gel with no band corresponding to the contrast medium peak.
“Shadow” artifact on an immunofixation gel caused by incomplete washing of monoclonal protein from the gel (lanes IgA, IgM, and lambda).
“Shadow” artifact on an immunofixation gel caused by incomplete washing of monoclonal protein from the gel (lanes IgA, IgM, and lambda).
(A) Densitometric scan from a concentrated urine specimen showing a monoclonal protein (1,000 mg/24 h) in the gamma region on urine PE. The patient had 0.7 g/dl of IgG lambda monoclonal protein (including C3 complement) in the beta globulin region on serum PE/IFE. (B) Urine IFE performed on the specimen used in panel A demonstrates a free lambda monoclonal protein.
(A) Densitometric scan from a concentrated urine specimen showing a monoclonal protein (1,000 mg/24 h) in the gamma region on urine PE. The patient had 0.7 g/dl of IgG lambda monoclonal protein (including C3 complement) in the beta globulin region on serum PE/IFE. (B) Urine IFE performed on the specimen used in panel A demonstrates a free lambda monoclonal protein.
IFE of a concentrated urine specimen demonstrating a dominant free kappa and a smaller amount of IgG kappa monoclonal protein.
IFE of a concentrated urine specimen demonstrating a dominant free kappa and a smaller amount of IgG kappa monoclonal protein.
IFE of a concentrated urine specimen demonstrating a free kappa and a free lambda monoclonal protein in a patient with IgA kappa and IgA lambda monoclonal proteins detected on serum IFE.
IFE of a concentrated urine specimen demonstrating a free kappa and a free lambda monoclonal protein in a patient with IgA kappa and IgA lambda monoclonal proteins detected on serum IFE.
Seven paired CSF (lanes 1 to 4 and 6 to 8) and serum (lanes 1′ to 4′ and 6′ to 8′) specimens along with two positive CSF controls (lanes 5 and 9) and two negative CSF controls (lanes 5′ and 9′) were subjected to IEF for the detection of oligoclonal bands. The patient specimens in lanes 1 and 1′ and 7 and 7′ reveal oligoclonal bands consistent with MS. The pattern for the patient in lanes 2 and 2′ are consistent with the presence of a monoclonal protein (posttranslational changes account for the multiple monoclonal protein bands seen with IEF). The patterns in lanes 3 and 3′ have faint bands appearing in both the CSF and serum specimens; this is nondiagnostic. The patterns in lanes 4 and 4′ and 8 and 8′ are normal.
Seven paired CSF (lanes 1 to 4 and 6 to 8) and serum (lanes 1′ to 4′ and 6′ to 8′) specimens along with two positive CSF controls (lanes 5 and 9) and two negative CSF controls (lanes 5′ and 9′) were subjected to IEF for the detection of oligoclonal bands. The patient specimens in lanes 1 and 1′ and 7 and 7′ reveal oligoclonal bands consistent with MS. The pattern for the patient in lanes 2 and 2′ are consistent with the presence of a monoclonal protein (posttranslational changes account for the multiple monoclonal protein bands seen with IEF). The patterns in lanes 3 and 3′ have faint bands appearing in both the CSF and serum specimens; this is nondiagnostic. The patterns in lanes 4 and 4′ and 8 and 8′ are normal.
Distribution of plasma cell proliferative disorders at the Mayo Clinic from 1960 to 2003 a
Distribution of plasma cell proliferative disorders at the Mayo Clinic from 1960 to 2003 a
Protocol for urine concentration used at Beaumont Laboratory
Protocol for urine concentration used at Beaumont Laboratory