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The Journal of Applied Laboratory... Nov 2020Laboratory methods for diagnosis and monitoring of monoclonal gammopathies have evolved to include serum and urine protein electrophoresis, immunofixation... (Review)
Review
BACKGROUND
Laboratory methods for diagnosis and monitoring of monoclonal gammopathies have evolved to include serum and urine protein electrophoresis, immunofixation electrophoresis, capillary zone electrophoresis, and immunosubtraction, serum-free light chain assay, mass spectrometry, and newly described QUIET.
CONTENT
This review presents a critical appraisal of the test methods and reporting practices for the findings generated by the tests for monoclonal gammopathies. Recommendations for desirable practices to optimize test selection and provide value-added reports are presented. The shortcomings of the serum-free light chain assay are highlighted, and new assays for measuring monoclonal serum free light chains are addressed.
SUMMARY
The various assays for screening, diagnosis, and monitoring of monoclonal gammopathies should be used in an algorithmic approach to avoid unnecessary testing. Reporting of the test results should be tailored to the clinical context of each individual patient to add value. Caution is urged in the interpretation of results of serum-free light chain assay, kappa/lambda ratio, and myeloma defining conditions. The distortions in serum-free light chain assay and development of oligoclonal bands in patients' status post hematopoietic stem cell transplants is emphasized and the need to note the location of original monoclonal Ig is stressed. The need for developing criteria that consider the differences in the biology of kappa and lambda light chain associated lesions is stressed. A new method of measuring monoclonal serum-free light chains is introduced. Reference is also made to a newly defined entity of light chain predominant intact immunoglobulin monoclonal gammopathy. The utility of urine testing in the diagnosis and monitoring of light chain only lesions is emphasized.
Topics: Electrophoresis; Humans; Immunoelectrophoresis; Immunoglobulin Light Chains; Immunoglobulin lambda-Chains; Paraproteinemias
PubMed: 33150391
DOI: 10.1093/jalm/jfaa153 -
Veterinary Clinical Pathology Dec 2022Hyperglobulinemia is reported in 26% of canine chronic B-cell lymphocytic leukemia (B-CLL) cases. However, few cases have been characterized by protein electrophoresis...
BACKGROUND
Hyperglobulinemia is reported in 26% of canine chronic B-cell lymphocytic leukemia (B-CLL) cases. However, few cases have been characterized by protein electrophoresis and immunofixation (IF), and the incidence of a monoclonal protein (M-protein) is unknown using these techniques.
OBJECTIVE
To characterize and determine the proportion of canine B-CLL cases with an M-protein using plasma protein electrophoresis (PPE), routine and free light chain (fLC) IF, and to assess if productive B-CLL cases express MUM1/IRF4 by cell tube block (CTB).
METHODS
PPE, routine (targeting IgG, IgA, IgM, IgG4, and light chain) and fLC IF were performed using 48 dog B-CLL plasma samples from patients diagnosed via peripheral blood flow cytometry. CTB was performed on a separate cohort of 15 patients.
RESULTS
Hyperproteinemia (>7.5 g/dL) was present in 17/48 cases (35%). An M-protein was detected in 32/48 cases (67%). Of these, 19/32 cases (59%) had only complete (monoclonal heavy and light chain) M-proteins detected, 10/32 cases (31%) had both complete and fLC M-proteins detected, and 3/32 cases (9%) had only an fLC M-protein detected. IgM was the most common clonal immunoglobulin isotype detected (23 cases). CD21 cell counts were higher in cases with detectable M-protein. Plasma fLC IF suggested β-γ region interference, likely caused by clotting proteins. All B-CLL cases consistently expressed PAX5 and did not express MUM1/IRF4.
CONCLUSIONS
Most B-CLL cases had an M-protein and were not hyperproteinemic. Most cases with paraproteins had a complete IgM monoclonal gammopathy; a subset had documented fLCs. The prognostic significance of heavy and fLC presence should be evaluated.
Topics: Dogs; Animals; Leukemia, Lymphocytic, Chronic, B-Cell; Immunoglobulin Light Chains; Immunoelectrophoresis; Paraproteinemias; Immunoglobulin M; Dog Diseases
PubMed: 35883213
DOI: 10.1111/vcp.13156 -
Archives of Pathology & Laboratory... Feb 1999A wide variety of techniques are available for the screening, characterization, and quantification of monoclonal proteins. These techniques vary in regard to the... (Review)
Review
A wide variety of techniques are available for the screening, characterization, and quantification of monoclonal proteins. These techniques vary in regard to the expense, skill and intensity of labor involved, and sensitivity for detection of low levels of monoclonal proteins or of those with unusual migration. Detection of monoclonal proteins requires the use of high-resolution electrophoresis (either gel-based or capillary) and immunofixation (or immunosubtraction). Immunoelectrophoresis is not recommended. Urine for detection of monoclonal free light chains should be from 24-hour samples, and the aliquot should be concentrated at least 100-fold prior to electrophoresis and immunofixation. Dipstick and sulfosalicylic acid techniques are not sensitive enough to detect small quantities of monoclonal free light chains and should not be used as screening tests for this purpose.
Topics: Electrophoresis; Humans; Immunoelectrophoresis; Immunoglobulins; Laboratories, Hospital; Paraproteinemias; Quality Assurance, Health Care
PubMed: 10050785
DOI: 10.5858/1999-123-0126-PFTEOM -
Annales de Biologie Clinique 2004
Topics: Bias; Humans; Immunoassay; Immunoelectrophoresis; Paraproteinemias; Patient Selection; Practice Guidelines as Topic; Reproducibility of Results
PubMed: 15297250
DOI: No ID Found -
Journal of Immunological Methods 1982Invented 20 years age, crossed immunoelectrophoresis (X-IEP) today is a technique of unusual power and myriad application. It combines very high resolution with... (Review)
Review
Invented 20 years age, crossed immunoelectrophoresis (X-IEP) today is a technique of unusual power and myriad application. It combines very high resolution with exquisite specificity by alloying 2-dimensional electrophoresis with immunoprecipitation for symbiotic new potentialities. The consequent matchless quantitative/qualitative capabilities of X-IEP for analyzing antigens in complex mixtures, particularly by their idiomatic internal comparison, are still not widely recognized. Because of this and the supposed complications of its use and interpretation, X-IEP is more rarely used than it should be. This essay discusses contemporary X-IEP with the particular aims of demonstrating that it is not difficult to use and of explaining with selected examples why it is peculiarly powerful for analyzing antigen mixtures like the body fluids, tissue and cell extracts, and microbial homogenates.
Topics: Analysis of Variance; Animals; Antigens; Antigens, Bacterial; Blood Proteins; Chemical Phenomena; Chemistry; Cross Reactions; Enzymes; Female; Humans; Hypergammaglobulinemia; Immune Sera; Immunoelectrophoresis; Immunoelectrophoresis, Two-Dimensional; Immunoglobulin G; Male; Mice; Mice, Inbred A; Rabbits; Respiratory Distress Syndrome; alpha 1-Antitrypsin
PubMed: 7045231
DOI: 10.1016/0022-1759(82)90218-6 -
Archives of Pathology & Laboratory... Feb 1999The first test for recognition of monoclonal gammopathies should be serum protein electrophoresis with high-resolution agarose gel. Serum protein electrophoresis should... (Review)
Review
The first test for recognition of monoclonal gammopathies should be serum protein electrophoresis with high-resolution agarose gel. Serum protein electrophoresis should be performed whenever multiple myeloma, Waldenström's macroglobulinemia, primary amyloidosis, or a related disorder is suspected. Immunofixation is critical for the differentiation of a monoclonal from a polyclonal increase in immunoglobulins. Quantitation of immunoglobulins should be performed with a rate nephelometer. The viscosity of serum should be measured if the patient has signs or symptoms of hyperviscosity syndrome. A 24-hour urine specimen should be obtained for determination of the total amount of protein excreted each day. Immunofixation of the urine should be performed on every patient who has an M-protein level greater than 1.5 g/dL (15 g/L) in the serum or in whom multiple myeloma, Waldenström's macroglobulinemia, primary amyloidosis, or a related disorder is suspected.
Topics: Electrophoresis; Humans; Immunoelectrophoresis; Laboratories, Hospital; Paraproteinemias
PubMed: 10050783
DOI: 10.5858/1999-123-0114-SOTFMG -
European Journal of Biochemistry Aug 1979Whole human platelets and platelet membranes have been solubilized in 1% Triton X-100, and the solubilized proteins examined by crossed immunoelectrophoresis using...
Characterization of human platelet proteins solubilized with Triton X-100 and examined by crossed immunoelectrophoresis. Reference patterns of extracts from whole platelets and isolated membranes.
Whole human platelets and platelet membranes have been solubilized in 1% Triton X-100, and the solubilized proteins examined by crossed immunoelectrophoresis using rabbit antibodies raised against either whole platelets or isolated membranes. 90% of the platelet proteins were solubilized by this extraction. About twenty immunoprecipitates were observed using the extracts obtained from whole platelets, whereas normally eight immunoprecipitates were seen with extracts from isolated membranes. Albumin, factor VIII and fibrinogen were identified with monospecific antibodies. Correlation of the patterns obtained for platelets or membranes was obtained by addition experiments, by crossed-line immunoelectrophoresis and by crossed immunoelectrophoresis of a mixture of extracts from unlabeled whole platelets and membranes isolated from platelets labeled by lactoperoxidase-catalyzed 125I iodination. Four sialoglycoproteins were identified by their reduced electrophoretic migration after neuraminidase treatment, and six proteins interacted with various lectins, indicating them to be glycosylated. Seven amphiphilic proteins were identified by charge-shift crossed immunoelectrophoresis, and nine by crossed hydrophobic interaction immunoelectrophoresis with phenyl-Sepharose. The topographical arrangement of the membrane proteins was examined with lactoperoxidase-catalyzed 125I-labeled platelets as antigens, and by antibodies absorbed with a suspension of whole platelets. Four and six radioactively labeled precipitates could be identified using the platelet and membrane extracts, respectively, indicating them to be exposed at the outer platelet surface. This was confirmed by the use of antibodies absorbed with intact platelets.
Topics: Blood Platelets; Blood Proteins; Cell Membrane; Humans; Immunoelectrophoresis, Two-Dimensional; Lectins; Membrane Proteins; Polyethylene Glycols; Solubility
PubMed: 488120
DOI: 10.1111/j.1432-1033.1979.tb13225.x -
Canadian Journal of Veterinary Research... Jul 1988Serum from 28 clinically healthy cats was subjected to agarose gel electrophoresis and the migration distance relative to albumin was determined. The reference values...
Serum from 28 clinically healthy cats was subjected to agarose gel electrophoresis and the migration distance relative to albumin was determined. The reference values for the relative and absolute concentrations of each protein fraction were determined and compared to previous reports. The immunoelectrophoretic, crossed immunoelectrophoretic and crossed line immunoelectrophoretic pattern, of a pooled sample of serum from clinically normal cats was determined. The cross-reactivity between goat and/or rabbit monospecific antisera to human proteins and feline serum was determined using immunoelectrophoresis and crossed-line absorption immunoelectrophoresis. Feline alpha-2-macroglobulin, haptoglobin, B1C-globulin, IgG, albumin and ceruloplasmin cross reacted strongly with the monospecific antisera. Alpha-2-macroglobulin migrated anodal to haptoglobin. Lipoproteins and ceruloplasmin were studied using staining procedures described in man. Feline transferrin was precipitated with Rivanol.
Topics: Animals; Blood Proteins; Cats; Ceruloplasmin; Cross Reactions; Densitometry; Electrophoresis, Agar Gel; Female; Immunoelectrophoresis; Immunoelectrophoresis, Two-Dimensional; Lipoproteins; Male; Reference Values; Staining and Labeling; Transferrin
PubMed: 2458817
DOI: No ID Found -
Blood Aug 2022
Topics: Antibodies, Monoclonal; COVID-19; Electrophoresis; Humans; Immunoelectrophoresis; Paraproteinemias
PubMed: 35925640
DOI: 10.1182/blood.2022016877 -
Gut May 1985The proteins of 46 human bile specimens, collected by several different routes have been studied by crossed immunoelectrophoresis, by rocket immunoelectrophoresis and by...
The proteins of 46 human bile specimens, collected by several different routes have been studied by crossed immunoelectrophoresis, by rocket immunoelectrophoresis and by radioimmunoassay. The results were analysed by plotting the variation in the bile: plasma ratio of particular proteins against molecular weight and by examination of the correlation between the concentrations of different proteins in the biles of different patients. Our results show that the majority of human bile proteins derive from plasma although bile specific proteins are always present. The majority of plasma proteins appear to enter bile by a 'sieving' mechanism which results in an inverse relationship between the bile: plasma ratio and the molecular weight. In addition there was a very high degree of correlation between the biliary concentrations of alpha 2-macroglobulin, IgG, haptoglobin, haemopexin, albumin, prealbumin, and orosomucoid. A number of other proteins namely thyroxine binding globulin, GC globulin and alpha 2HS-glycoprotein appeared in bile at concentrations greater than those expected if entry is by the sieving mechanism. These three proteins, however, are of rather low molecular weight and the reason for the lack of correlation appears to be individual variation in the 'pore size', presumably reflecting variation in the porosity of tight junction between hepatocytes. Although the majority of human bile proteins would appear to enter bile by a molecular weight-dependent pathway, four proteins, namely secretory IgA, IgM, haemoglobin and caeruloplasmin, showed significant deviation from the predicted relationship and probably enter bile at least partly by transport across cells. The concentration of beta 2-glycoprotein I was also much greater than expected from its molecular weight. The reason for this is not yet clear but may well reflect a very efficient and specific transport mechanism.
Topics: Bile; Blood Proteins; Humans; Immunoelectrophoresis; Immunoelectrophoresis, Two-Dimensional; Molecular Weight; Proteins; Radioimmunoassay
PubMed: 3996941
DOI: 10.1136/gut.26.5.500