Polyclonal antibodies against kappa light chain are used to diagnose diseases producing free light chain. The kappa and lambda light chains are products of immunoglobulin synthesis and released into the circulation in minor amounts such as serum, cerebrospinal fluid, urine and synovial fluid in normal condition.
The purpose of this study was the production and purification of polyclonal immunoglobulin G (IgG) against human kappa light chains.
In this study, early human IgG was purified by ion-exchange chromatography, reduced with Dithiothreitol and heavy and light chains were separated with size-exclusion chromatography.
Afterward, affinity chromatography with protein L Sepharose at pH 2.00 was displayed to be a dominant condition for the separation and purification of the kappa light chain of immunoglobulins from human serum.
Eventually, the rabbit was immunized by human kappa light chains. The rabbit IgG was purified and labeled with horseradish peroxidase (HRP).
Direct enzyme-linked immunosorbent assay was planned to determine the titer of HRP conjugated rabbit IgG against the human kappa light chain.
The optimum titer of anti-kappa IgG was 1:16000. At the result, purified polyclonal anti-kappa is useful tool in biomedical and biochemical researches and diagnostic kits.
Membranous Nephropathy with Lambda Light Chain Restriction: A Rare Form with Serum Negative and Tissue Positive PLA2R Ab
Membranous nephropathy (MN) is the most common cause of nephrotic syndrome in adults.
Subepithelial polyclonal immunoglobulin deposits and >70% M-type phospholipase A2 receptor antibody positivity are typical findings in idiopathic MN.
A 58-year-old female patient was admitted with clinical presentation of nephrotic syndrome. Autoimmune diseases, infections, and malignancies were ruled out after clinical and laboratory evaluations.
Diagnostic work-up revealed serum PLA2R antibody negativity and diffuse thickening of glomerular capillary wall on biopsy, while glomerular capillary wall IgG, C3, and Lambda monotypic light chain deposition and PLA2R1 positivity were detected by immunofluorescence and immunohistochemical examination, respectively. Following prednisolone treatment, creatinine and proteinuria were markedly regressed. The MN cases with a light chain deposits are rare and experience regarding their treatment are insufficient.
Can Antinuclear Antibodies (ANA) be Monoclonal?
Background: Nuclear staining by immunofluorescence in a kidney biopsy is often seen in patients with positive antinuclear antibodies (ANA) in the serum.
These ANA are usually polyclonal, but herein we report 9 cases with an unusual finding of monoclonal nuclear staining by immunofluorescence on kidney biopsy.
Case Presentation.
Nine cases with predominant stain for kappa or lambda light chain were identified by searching the renal pathology laboratory database for the past 10 years.
All cases had positive stain for only kappa (six cases) or lambda (three cases) light chain in the nuclei.
Eight out of nine cases had positive nuclear IgG stain, and one case had positive nuclear IgA stain. Among cases with positive nuclear IgG staining, six cases were positive for IgGIgGIgGIgG nuclear stain, who had testing for ANA, had positive ANA. Patients with positive IgGIgGlight chain stain in the nuclei had IgG lambda monoclonal gammopathy.
Immune subtraction for improved resolution in serum protein immunofixation electrophoresis and antibody isotype determination in a patient with autoantibody
Heavy chain isotypes of low level monoclonal immunoglobulins are sometimes obscured in serum immunofixation electrophoresis (SIFE) by a heavy background of polyclonal immunoglobulins.
However, accurate determination of the heavy chain isotype is essential for a complete diagnosis, as isotype determination of autoantibodies may have relevance in determining therapeutic procedures. Immune subtraction (IS) was employed in a patient with neuropathy and GD1a autoantibody.
IS allowed identification of the cognate heavy chain related to a lambda light chain restriction noted on initial SIFE as well as isotype determination of the autoantibody.
Antisera specific to individual heavy and light chains were used for depletion of specific immunoglobulin types.
Depletion of kappa light chain associated immunoglobulins allowed unequivocal determination of the isotype of lambda light chain-associated low level monoclonal band to be IgG Lambda.
Selective depletion of kappa, lambda, gamma and mu heavy chain immunoglobulins was employed to determine IgG Kappa isotype of the auto-antibody.
Diagnostic performance of routine electrophoresis and immunofixation for the detection of immunoglobulin paraproteins (M-Proteins) in dogs with multiple myeloma and related disorders: Part 2-Toward improved diagnostic performance
Background: The diagnostic performance of routine electrophoresis (agarose gel electrophoresis [AGE] and capillary zone electrophoresis [CZE]) and species-specific immunofixation (IF) for the detection of immunoglobulin paraproteins (M-proteins) and diagnosis of secretory myeloma-related disorders (sMRD) can be improved.
Available canine IF targets were IgG-FC, IgA, IgM, light chain (LC), IgGantibodies.
Objective: We aimed to review specific features associated with the presence of M-proteins in canine serum samples and the common features causing inaccurate reporting of M-proteins to improve the diagnostic performance of routine electrophoresis and IF for the detection of M-proteins.
<strong class=”sub-title”> Methods: </strong> Features found in AGE, CZE, routine IF, <em>IgG</em>4 IF, and fLC IF of 100 canine serum samples from Part 1 of this study were evaluated by simple and multivariate logistic regression to identify factors associated with the presence of M-proteins.
Cases falsely called negative or positive for M-proteins were reviewed to identify the common features that could be used to increase the diagnostic performance of SPE and IF for M-protein detection.
Results: The presence of hypogammaglobulinemia or any peak taller than albumin was associated with an M-protein. Total protein concentrations, globulin concentrations, or peaks wider than albumin were not associated with an M-protein.
Free LC sMRD cases were not diagnosed by SPE and routine IF.
Cases with infectious and inflammatory etiologies had a restricted polyclonal gammopathy with multiple γ-globulin restrictions resulting in some false-positive results. SPE combined with all available IF results and the specific features identified in this study had an estimated sensitivity of 95.1% and specificity of 81.4%.
Conclusions: The identified criteria of this study increase the diagnostic performance of the electrophoretic evaluation for M-proteins.
Keywords: Canis lupis familiaris; biclonal gammopathy; canine; ehrlichia; immunotyping; monoclonal gammopathy.
Immunofluorescence Staining for Immunoglobulin Heavy Chain/LightChain on Kidney Biopsies is a Valuable Ancillary Technique For the Diagnosis of Monoclonal Gammopathy-Associated Kidney Diseases
Heavy chain/light chain (HLC) antibodies target conformational epitopes at the junctions of the heavy chain and light chain constant regions (CH1 and CL) of serum IgGκ, IgGλ, IgAκ, IgAλ, IgMκ, and IgMλ to provide quantitation of intact HLC pairs.
Here, we developed an HLC tissue immunofluorescence protocol to test if it can complement conventional immunofluorescence in the diagnosis of monoclonal gammopathy-associated kidney diseases.
HLC immunofluorescence was performed on archived frozen tissue of 104 kidney biopsies.
The sensitivity and specificity of HLC immunofluorescence was confirmed by testing cases of lupus nephritis, other polyclonal immunoglobulin nephropathies, and light chain nephropathies (light chain amyloidosis and deposition disease).
Testing of ten cases of the IgG variant of proliferative glomerulonephritis with monoclonal immunoglobulin deposits excluded monoclonal deposits in two by revealing positivity for IgGκ and IgGλ. Testing of 12 cases of monotypic IgA nephropathy excluded monoclonal deposits in six by revealing staining for IgAκ and IgAλ.
Testing of six cases of monotypic fibrillary glomerulonephritis excluded monoclonal deposits in three by revealing positivity for IgGκ and IgGλ.
None of 14 cases of glomerulonephritis in which HLC immunofluorescence unmasked polytypic deposits were associated with a serum or urine monoclonal immunoglobulins matching the conventional immunofluorescence results.
igg light chain Polyclonal Antibody |
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| 42162 | SAB | 100μg | 390 EUR |
igg light chain Polyclonal Antibody |
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| 42162-100ul | SAB | 100ul | 399.6 EUR |
igg Light Chain Polyclonal Antibody |
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| A57596 | EpiGentek |
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igg Light Chain Polyclonal Antibody |
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| A57793 | EpiGentek |
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igg light chain Polyclonal Antibody |
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| MBS9415682-01mg | MyBiosource | 0.1mg | 350 EUR |
igg light chain Polyclonal Antibody |
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| MBS9415682-5x01mg | MyBiosource | 5x0.1mg | 1420 EUR |
igg light chain Polyclonal Conjugated Antibody |
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| C42162 | SAB | 100ul | 476.4 EUR |
IgG Light Chain Polyclonal Conjugated Antibody |
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| MBS9450593-01mLAF350 | MyBiosource | 0.1mL(AF350) | 480 EUR |
IgG Light Chain Polyclonal Conjugated Antibody |
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| MBS9450593-01mLAF405 | MyBiosource | 0.1mL(AF405) | 480 EUR |
IgG Light Chain Polyclonal Conjugated Antibody |
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| MBS9450593-01mLAF488 | MyBiosource | 0.1mL(AF488) | 480 EUR |
HLC immunofluorescence outperformed paraffin immunofluorescence and IgG subclass staining in 10/13 (77%) of cases.
Testing of 18 cases of cryoglobulinemic glomerulonephritis showed better correlation with serum cryoprecipitate immunofixation than conventional immunofluorescence with regards to the type of cryoglobulin in 47% of cases.
Thus, HLC immunofluorescence is a valuable ancillary technique in kidney pathology for the diagnosis of monoclonal gammopathy-associated nephropathies, and could be utilized to confirm or exclude the monoclonal nature of deposits.