Assessment of Intrathecal Free Light Chain Synthesis: Comparison of Different Quantitative Methods with the Detection of Oligoclonal Free Light Chains by Isoelectric Focusing and Affinity-Mediated Immunoblotting
We aimed to compare various methods for free light chain (fLC) quantitation in cerebrospinal fluid (CSF) and serum and to determine whether quantitative CSF measurements could reliably predict intrathecal fLC synthesis. In addition, we wished to determine the relationship between free kappa and free...
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| creator | Zeman, David Kušnierová, Pavlína Švagera, Zdeněk Všianský, František Byrtusová, Monika Hradílek, Pavel Kurková, Barbora Zapletalová, Olga Bartoš, Vladimír |
| description | We aimed to compare various methods for free light chain (fLC) quantitation in cerebrospinal fluid (CSF) and serum and to determine whether quantitative CSF measurements could reliably predict intrathecal fLC synthesis. In addition, we wished to determine the relationship between free kappa and free lambda light chain concentrations in CSF and serum in various disease groups.
We analysed 166 paired CSF and serum samples by at least one of the following methods: turbidimetry (Freelite™, SPAPLUS), nephelometry (N Latex FLC™, BN ProSpec), and two different (commercially available and in-house developed) sandwich ELISAs. The results were compared with oligoclonal fLC detected by affinity-mediated immunoblotting after isoelectric focusing.
Although the correlations between quantitative methods were good, both proportional and systematic differences were discerned. However, no major differences were observed in the prediction of positive oligoclonal fLC test. Surprisingly, CSF free kappa/free lambda light chain ratios were lower than those in serum in about 75% of samples with negative oligoclonal fLC test. In about a half of patients with multiple sclerosis and clinically isolated syndrome, profoundly increased free kappa/free lambda light chain ratios were found in the CSF.
Our results show that using appropriate method-specific cut-offs, different methods of CSF fLC quantitation can be used for the prediction of intrathecal fLC synthesis. The reason for unexpectedly low free kappa/free lambda light chain ratios in normal CSFs remains to be elucidated. Whereas CSF free kappa light chain concentration is increased in most patients with multiple sclerosis and clinically isolated syndrome, CSF free lambda light chain values show large interindividual variability in these patients and should be investigated further for possible immunopathological and prognostic significance. |
| doi_str_mv | 10.1371/journal.pone.0166556 |
| format | Article |
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We analysed 166 paired CSF and serum samples by at least one of the following methods: turbidimetry (Freelite™, SPAPLUS), nephelometry (N Latex FLC™, BN ProSpec), and two different (commercially available and in-house developed) sandwich ELISAs. The results were compared with oligoclonal fLC detected by affinity-mediated immunoblotting after isoelectric focusing.
Although the correlations between quantitative methods were good, both proportional and systematic differences were discerned. However, no major differences were observed in the prediction of positive oligoclonal fLC test. Surprisingly, CSF free kappa/free lambda light chain ratios were lower than those in serum in about 75% of samples with negative oligoclonal fLC test. In about a half of patients with multiple sclerosis and clinically isolated syndrome, profoundly increased free kappa/free lambda light chain ratios were found in the CSF.
Our results show that using appropriate method-specific cut-offs, different methods of CSF fLC quantitation can be used for the prediction of intrathecal fLC synthesis. The reason for unexpectedly low free kappa/free lambda light chain ratios in normal CSFs remains to be elucidated. Whereas CSF free kappa light chain concentration is increased in most patients with multiple sclerosis and clinically isolated syndrome, CSF free lambda light chain values show large interindividual variability in these patients and should be investigated further for possible immunopathological and prognostic significance.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0166556</identifier><identifier>PMID: 27846293</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Affinity ; Biology and Life Sciences ; Case-Control Studies ; Cerebrospinal fluid ; Chains ; Demyelinating Diseases - blood ; Demyelinating Diseases - cerebrospinal fluid ; Demyelinating Diseases - diagnosis ; Enzyme-Linked Immunosorbent Assay - instrumentation ; Enzyme-Linked Immunosorbent Assay - methods ; Fluids ; Focusing ; Humans ; Immunoblotting ; Immunoblotting - instrumentation ; Immunoblotting - methods ; Immunoglobulin kappa-Chains - biosynthesis ; Immunoglobulin kappa-Chains - blood ; Immunoglobulin kappa-Chains - cerebrospinal fluid ; Immunoglobulin lambda-Chains - biosynthesis ; Immunoglobulin lambda-Chains - blood ; Immunoglobulin lambda-Chains - cerebrospinal fluid ; Immunoglobulins ; Isoelectric focusing ; Isoelectric Focusing - instrumentation ; Isoelectric Focusing - methods ; Laboratories ; Latex ; Light ; Light chains ; Medical diagnosis ; Medicine and Health Sciences ; Methods ; Multiple sclerosis ; Multiple Sclerosis - blood ; Multiple Sclerosis - cerebrospinal fluid ; Multiple Sclerosis - diagnosis ; Nephelometry ; Nephelometry and Turbidimetry - instrumentation ; Nephelometry and Turbidimetry - methods ; Neurology ; Observer Variation ; Patients ; Quantitation ; Reproducibility of Results ; Research and Analysis Methods ; ROC Curve ; Studies ; Synthesis ; Turbidimetry ; Urine</subject><ispartof>PloS one, 2016-11, Vol.11 (11), p.e0166556-e0166556</ispartof><rights>COPYRIGHT 2016 Public Library of Science</rights><rights>2016 Zeman et al. This is an open access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2016 Zeman et al 2016 Zeman et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c725t-967c39c9ded35f3f8605fe807c5c2fba01d79762040a80d9d382f6426e77a9e73</citedby><cites>FETCH-LOGICAL-c725t-967c39c9ded35f3f8605fe807c5c2fba01d79762040a80d9d382f6426e77a9e73</cites><orcidid>0000-0001-6822-2572</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC5112955/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC5112955/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,314,723,776,780,860,881,2096,2915,23845,27901,27902,53766,53768,79343,79344</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/27846293$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Zeman, David</creatorcontrib><creatorcontrib>Kušnierová, Pavlína</creatorcontrib><creatorcontrib>Švagera, Zdeněk</creatorcontrib><creatorcontrib>Všianský, František</creatorcontrib><creatorcontrib>Byrtusová, Monika</creatorcontrib><creatorcontrib>Hradílek, Pavel</creatorcontrib><creatorcontrib>Kurková, Barbora</creatorcontrib><creatorcontrib>Zapletalová, Olga</creatorcontrib><creatorcontrib>Bartoš, Vladimír</creatorcontrib><title>Assessment of Intrathecal Free Light Chain Synthesis: Comparison of Different Quantitative Methods with the Detection of Oligoclonal Free Light Chains by Isoelectric Focusing and Affinity-Mediated Immunoblotting</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>We aimed to compare various methods for free light chain (fLC) quantitation in cerebrospinal fluid (CSF) and serum and to determine whether quantitative CSF measurements could reliably predict intrathecal fLC synthesis. In addition, we wished to determine the relationship between free kappa and free lambda light chain concentrations in CSF and serum in various disease groups.
We analysed 166 paired CSF and serum samples by at least one of the following methods: turbidimetry (Freelite™, SPAPLUS), nephelometry (N Latex FLC™, BN ProSpec), and two different (commercially available and in-house developed) sandwich ELISAs. The results were compared with oligoclonal fLC detected by affinity-mediated immunoblotting after isoelectric focusing.
Although the correlations between quantitative methods were good, both proportional and systematic differences were discerned. However, no major differences were observed in the prediction of positive oligoclonal fLC test. Surprisingly, CSF free kappa/free lambda light chain ratios were lower than those in serum in about 75% of samples with negative oligoclonal fLC test. In about a half of patients with multiple sclerosis and clinically isolated syndrome, profoundly increased free kappa/free lambda light chain ratios were found in the CSF.
Our results show that using appropriate method-specific cut-offs, different methods of CSF fLC quantitation can be used for the prediction of intrathecal fLC synthesis. The reason for unexpectedly low free kappa/free lambda light chain ratios in normal CSFs remains to be elucidated. Whereas CSF free kappa light chain concentration is increased in most patients with multiple sclerosis and clinically isolated syndrome, CSF free lambda light chain values show large interindividual variability in these patients and should be investigated further for possible immunopathological and prognostic significance.</description><subject>Affinity</subject><subject>Biology and Life Sciences</subject><subject>Case-Control Studies</subject><subject>Cerebrospinal fluid</subject><subject>Chains</subject><subject>Demyelinating Diseases - blood</subject><subject>Demyelinating Diseases - cerebrospinal fluid</subject><subject>Demyelinating Diseases - diagnosis</subject><subject>Enzyme-Linked Immunosorbent Assay - instrumentation</subject><subject>Enzyme-Linked Immunosorbent Assay - methods</subject><subject>Fluids</subject><subject>Focusing</subject><subject>Humans</subject><subject>Immunoblotting</subject><subject>Immunoblotting - instrumentation</subject><subject>Immunoblotting - methods</subject><subject>Immunoglobulin kappa-Chains - biosynthesis</subject><subject>Immunoglobulin kappa-Chains - blood</subject><subject>Immunoglobulin kappa-Chains - cerebrospinal fluid</subject><subject>Immunoglobulin lambda-Chains - biosynthesis</subject><subject>Immunoglobulin lambda-Chains - blood</subject><subject>Immunoglobulin lambda-Chains - cerebrospinal fluid</subject><subject>Immunoglobulins</subject><subject>Isoelectric focusing</subject><subject>Isoelectric Focusing - instrumentation</subject><subject>Isoelectric Focusing - methods</subject><subject>Laboratories</subject><subject>Latex</subject><subject>Light</subject><subject>Light chains</subject><subject>Medical diagnosis</subject><subject>Medicine and Health Sciences</subject><subject>Methods</subject><subject>Multiple sclerosis</subject><subject>Multiple Sclerosis - blood</subject><subject>Multiple Sclerosis - cerebrospinal fluid</subject><subject>Multiple Sclerosis - diagnosis</subject><subject>Nephelometry</subject><subject>Nephelometry and Turbidimetry - instrumentation</subject><subject>Nephelometry and Turbidimetry - methods</subject><subject>Neurology</subject><subject>Observer Variation</subject><subject>Patients</subject><subject>Quantitation</subject><subject>Reproducibility of Results</subject><subject>Research and Analysis Methods</subject><subject>ROC Curve</subject><subject>Studies</subject><subject>Synthesis</subject><subject>Turbidimetry</subject><subject>Urine</subject><issn>1932-6203</issn><issn>1932-6203</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2016</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><sourceid>BENPR</sourceid><sourceid>DOA</sourceid><recordid>eNqNk9FuFCEUhidGY7X6BkZJTIxe7DoMAzN4YbLZWt2kTaNVbwkLhx2aGVgHprrP6QvJdLdN1_Si4QIC3_8fOIeTZS9wPsWkwu8v_NA72U7X3sE0x4xRyh5kTzAnxYQVOXl4a32QPQ3hIs8pqRl7nB0UVV2ygpMn2d9ZCBBCBy4ib9DCxV7GBpRs0XEPgE7sqolo3kjr0PnGpaNgwwc0991a9jZ4N6qOrDHQjxZfB-mijTLaS0CnEBuvA_ptY4OSEh1BBBXtVnTW2pVXrXd3hApouUGL4KFNfG8VOvZqCNatkHQazYyxzsbN5BS0lRE0WnTd4Pyy9TEm6Fn2yMg2wPPdfJj9OP70ff5lcnL2eTGfnUxUVdA44axShCuuQRNqiKlZTg3UeaWoKsxS5lhXvErpK3NZ55prUheGlQWDqpIcKnKYvdr6rlsfxK4eQeC6xJhWOS8TsdgS2ssLse5tJ_uN8NKKqw3fr4Tso1UtCGVorUotOS2SEBjHoKkmmi0pB1Po5PVxF21YdqAVjKVq90z3T5xtxMpfCopxwSlNBm93Br3_NUCIorNBQdtKB364ujcrMStJcQ-U8IowUozo6__QuxOxo1YyvdU649MV1WgqZmWFa0aSY6Kmd1BpaOisSv_c2LS_J3i3J0hMhD9xJYcQxOL82_3Zs5_77JtbbAOyjU3w7TD-3bAPlltQ9T6EHsxNPXAuxja9zoYY21Ts2jTJXt6u5Y3oui_JP7AtO8s</recordid><startdate>20161115</startdate><enddate>20161115</enddate><creator>Zeman, David</creator><creator>Kušnierová, Pavlína</creator><creator>Švagera, Zdeněk</creator><creator>Všianský, František</creator><creator>Byrtusová, Monika</creator><creator>Hradílek, Pavel</creator><creator>Kurková, Barbora</creator><creator>Zapletalová, Olga</creator><creator>Bartoš, Vladimír</creator><general>Public Library of Science</general><general>Public Library of Science (PLoS)</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>IOV</scope><scope>ISR</scope><scope>3V.</scope><scope>7QG</scope><scope>7QL</scope><scope>7QO</scope><scope>7RV</scope><scope>7SN</scope><scope>7SS</scope><scope>7T5</scope><scope>7TG</scope><scope>7TM</scope><scope>7U9</scope><scope>7X2</scope><scope>7X7</scope><scope>7XB</scope><scope>88E</scope><scope>8AO</scope><scope>8C1</scope><scope>8FD</scope><scope>8FE</scope><scope>8FG</scope><scope>8FH</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABJCF</scope><scope>ABUWG</scope><scope>AEUYN</scope><scope>AFKRA</scope><scope>ARAPS</scope><scope>ATCPS</scope><scope>AZQEC</scope><scope>BBNVY</scope><scope>BENPR</scope><scope>BGLVJ</scope><scope>BHPHI</scope><scope>C1K</scope><scope>CCPQU</scope><scope>D1I</scope><scope>DWQXO</scope><scope>FR3</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>GNUQQ</scope><scope>H94</scope><scope>HCIFZ</scope><scope>K9.</scope><scope>KB.</scope><scope>KB0</scope><scope>KL.</scope><scope>L6V</scope><scope>LK8</scope><scope>M0K</scope><scope>M0S</scope><scope>M1P</scope><scope>M7N</scope><scope>M7P</scope><scope>M7S</scope><scope>NAPCQ</scope><scope>P5Z</scope><scope>P62</scope><scope>P64</scope><scope>PATMY</scope><scope>PDBOC</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>PTHSS</scope><scope>PYCSY</scope><scope>RC3</scope><scope>7X8</scope><scope>5PM</scope><scope>DOA</scope><orcidid>https://orcid.org/0000-0001-6822-2572</orcidid></search><sort><creationdate>20161115</creationdate><title>Assessment of Intrathecal Free Light Chain Synthesis: Comparison of Different Quantitative Methods with the Detection of Oligoclonal Free Light Chains by Isoelectric Focusing and Affinity-Mediated Immunoblotting</title><author>Zeman, David ; Kušnierová, Pavlína ; Švagera, Zdeněk ; Všianský, František ; Byrtusová, Monika ; Hradílek, Pavel ; Kurková, Barbora ; Zapletalová, Olga ; Bartoš, Vladimír</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c725t-967c39c9ded35f3f8605fe807c5c2fba01d79762040a80d9d382f6426e77a9e73</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2016</creationdate><topic>Affinity</topic><topic>Biology and Life Sciences</topic><topic>Case-Control Studies</topic><topic>Cerebrospinal fluid</topic><topic>Chains</topic><topic>Demyelinating Diseases - 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Academic</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>PloS one</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Zeman, David</au><au>Kušnierová, Pavlína</au><au>Švagera, Zdeněk</au><au>Všianský, František</au><au>Byrtusová, Monika</au><au>Hradílek, Pavel</au><au>Kurková, Barbora</au><au>Zapletalová, Olga</au><au>Bartoš, Vladimír</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Assessment of Intrathecal Free Light Chain Synthesis: Comparison of Different Quantitative Methods with the Detection of Oligoclonal Free Light Chains by Isoelectric Focusing and Affinity-Mediated Immunoblotting</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2016-11-15</date><risdate>2016</risdate><volume>11</volume><issue>11</issue><spage>e0166556</spage><epage>e0166556</epage><pages>e0166556-e0166556</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>We aimed to compare various methods for free light chain (fLC) quantitation in cerebrospinal fluid (CSF) and serum and to determine whether quantitative CSF measurements could reliably predict intrathecal fLC synthesis. In addition, we wished to determine the relationship between free kappa and free lambda light chain concentrations in CSF and serum in various disease groups.
We analysed 166 paired CSF and serum samples by at least one of the following methods: turbidimetry (Freelite™, SPAPLUS), nephelometry (N Latex FLC™, BN ProSpec), and two different (commercially available and in-house developed) sandwich ELISAs. The results were compared with oligoclonal fLC detected by affinity-mediated immunoblotting after isoelectric focusing.
Although the correlations between quantitative methods were good, both proportional and systematic differences were discerned. However, no major differences were observed in the prediction of positive oligoclonal fLC test. Surprisingly, CSF free kappa/free lambda light chain ratios were lower than those in serum in about 75% of samples with negative oligoclonal fLC test. In about a half of patients with multiple sclerosis and clinically isolated syndrome, profoundly increased free kappa/free lambda light chain ratios were found in the CSF.
Our results show that using appropriate method-specific cut-offs, different methods of CSF fLC quantitation can be used for the prediction of intrathecal fLC synthesis. The reason for unexpectedly low free kappa/free lambda light chain ratios in normal CSFs remains to be elucidated. Whereas CSF free kappa light chain concentration is increased in most patients with multiple sclerosis and clinically isolated syndrome, CSF free lambda light chain values show large interindividual variability in these patients and should be investigated further for possible immunopathological and prognostic significance.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>27846293</pmid><doi>10.1371/journal.pone.0166556</doi><tpages>e0166556</tpages><orcidid>https://orcid.org/0000-0001-6822-2572</orcidid><oa>free_for_read</oa></addata></record> |
| fulltext | fulltext |
| identifier | ISSN: 1932-6203 |
| ispartof | PloS one, 2016-11, Vol.11 (11), p.e0166556-e0166556 |
| issn | 1932-6203 1932-6203 |
| language | eng |
| recordid | cdi_plos_journals_1841157094 |
| source | MEDLINE; DOAJ Directory of Open Access Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; PubMed Central; Free Full-Text Journals in Chemistry; Public Library of Science (PLoS) |
| subjects | Affinity Biology and Life Sciences Case-Control Studies Cerebrospinal fluid Chains Demyelinating Diseases - blood Demyelinating Diseases - cerebrospinal fluid Demyelinating Diseases - diagnosis Enzyme-Linked Immunosorbent Assay - instrumentation Enzyme-Linked Immunosorbent Assay - methods Fluids Focusing Humans Immunoblotting Immunoblotting - instrumentation Immunoblotting - methods Immunoglobulin kappa-Chains - biosynthesis Immunoglobulin kappa-Chains - blood Immunoglobulin kappa-Chains - cerebrospinal fluid Immunoglobulin lambda-Chains - biosynthesis Immunoglobulin lambda-Chains - blood Immunoglobulin lambda-Chains - cerebrospinal fluid Immunoglobulins Isoelectric focusing Isoelectric Focusing - instrumentation Isoelectric Focusing - methods Laboratories Latex Light Light chains Medical diagnosis Medicine and Health Sciences Methods Multiple sclerosis Multiple Sclerosis - blood Multiple Sclerosis - cerebrospinal fluid Multiple Sclerosis - diagnosis Nephelometry Nephelometry and Turbidimetry - instrumentation Nephelometry and Turbidimetry - methods Neurology Observer Variation Patients Quantitation Reproducibility of Results Research and Analysis Methods ROC Curve Studies Synthesis Turbidimetry Urine |
| title | Assessment of Intrathecal Free Light Chain Synthesis: Comparison of Different Quantitative Methods with the Detection of Oligoclonal Free Light Chains by Isoelectric Focusing and Affinity-Mediated Immunoblotting |
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