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A method to obtain purified free light chain monomers and dimers from urine samples of patients with multiple myeloma

Antibody light chains are synthesized in excess by plasma cells, and this excess can be secreted into biological fluids as dimers or monomers in various proportions. Structural differences between monomers or dimers of free light chains (FLC) can affect their biological functions and possibly their...

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Autores principales: Caponi, Laura, Botti, Alice, Romiti, Nadia, Paolicchi, Aldo, Franzini, Maria
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer US 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9659499/
https://www.ncbi.nlm.nih.gov/pubmed/36103008
http://dx.doi.org/10.1007/s12026-022-09314-8
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author Caponi, Laura
Botti, Alice
Romiti, Nadia
Paolicchi, Aldo
Franzini, Maria
author_facet Caponi, Laura
Botti, Alice
Romiti, Nadia
Paolicchi, Aldo
Franzini, Maria
author_sort Caponi, Laura
collection PubMed
description Antibody light chains are synthesized in excess by plasma cells, and this excess can be secreted into biological fluids as dimers or monomers in various proportions. Structural differences between monomers or dimers of free light chains (FLC) can affect their biological functions and possibly their pathogenicity. They also may exhibit differential immune reactivity, perhaps explaining discrepant quantifications when measured by different immunoreagents. Having purified FLC monomers and dimers available can be useful for studying their properties. Here we propose a simple preparatory procedure to purify FLC monomers and dimers from urine samples of patients with plasma cell disorders. Two representative urine samples containing lambda or kappa FLC were loaded into a nonreducing sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE). The gel strips containing separate monomers and dimers were excised, electroeluted, and the FLC recovered. The FLC were recovered from SDS-PAGE gel in sufficient amounts to be quantified by UV and two automated nephelometric assays immunochemical. The procedure was found to be simple, reproducible, and with a high yield, thus offering the opportunity to compare different assays. Not all urine samples are suitable for this procedure, but this approach allows for the purification of FLC monomers and dimers from many selected urine samples which maintain their oligomeric organization.
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spelling pubmed-96594992022-11-15 A method to obtain purified free light chain monomers and dimers from urine samples of patients with multiple myeloma Caponi, Laura Botti, Alice Romiti, Nadia Paolicchi, Aldo Franzini, Maria Immunol Res Original Article Antibody light chains are synthesized in excess by plasma cells, and this excess can be secreted into biological fluids as dimers or monomers in various proportions. Structural differences between monomers or dimers of free light chains (FLC) can affect their biological functions and possibly their pathogenicity. They also may exhibit differential immune reactivity, perhaps explaining discrepant quantifications when measured by different immunoreagents. Having purified FLC monomers and dimers available can be useful for studying their properties. Here we propose a simple preparatory procedure to purify FLC monomers and dimers from urine samples of patients with plasma cell disorders. Two representative urine samples containing lambda or kappa FLC were loaded into a nonreducing sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE). The gel strips containing separate monomers and dimers were excised, electroeluted, and the FLC recovered. The FLC were recovered from SDS-PAGE gel in sufficient amounts to be quantified by UV and two automated nephelometric assays immunochemical. The procedure was found to be simple, reproducible, and with a high yield, thus offering the opportunity to compare different assays. Not all urine samples are suitable for this procedure, but this approach allows for the purification of FLC monomers and dimers from many selected urine samples which maintain their oligomeric organization. Springer US 2022-09-14 2022 /pmc/articles/PMC9659499/ /pubmed/36103008 http://dx.doi.org/10.1007/s12026-022-09314-8 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Original Article
Caponi, Laura
Botti, Alice
Romiti, Nadia
Paolicchi, Aldo
Franzini, Maria
A method to obtain purified free light chain monomers and dimers from urine samples of patients with multiple myeloma
title A method to obtain purified free light chain monomers and dimers from urine samples of patients with multiple myeloma
title_full A method to obtain purified free light chain monomers and dimers from urine samples of patients with multiple myeloma
title_fullStr A method to obtain purified free light chain monomers and dimers from urine samples of patients with multiple myeloma
title_full_unstemmed A method to obtain purified free light chain monomers and dimers from urine samples of patients with multiple myeloma
title_short A method to obtain purified free light chain monomers and dimers from urine samples of patients with multiple myeloma
title_sort method to obtain purified free light chain monomers and dimers from urine samples of patients with multiple myeloma
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9659499/
https://www.ncbi.nlm.nih.gov/pubmed/36103008
http://dx.doi.org/10.1007/s12026-022-09314-8
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