Detection of Bence-Jones Protein in Serum by Immunoblotting
To detect Bence-Jones protein (BJP) in serum we precipitated intact immunoglobulins (Ig) using polyethylene glycol (PEG) and subjected the BJP in solution to electrophoresis in agarose gel, followed by transfer to a polyvinylidene difluoride membrane, and immunoenzymatic staining (successively using...
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| Veröffentlicht in: | Annals of clinical biochemistry 1996-03, Vol.33 (2), p.132-138 |
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| creator | Backer, E T Brand, A |
| description | To detect Bence-Jones protein (BJP) in serum we precipitated intact immunoglobulins (Ig) using polyethylene glycol (PEG) and subjected the BJP in solution to electrophoresis in agarose gel, followed by transfer to a polyvinylidene difluoride membrane, and immunoenzymatic staining (successively using rabbit anti-human light/heavy chain of Ig, biotinylated swine anti-rabbit Ig, and alkaline phosphatase-conjugated streptavidin). Treatment with PEG effectively reduced background staining of polyclonal Ig in the immunoblots, although intact monoclonal Ig was not always completely removed. To compare the present method with immunoelectrophoresis (IEP), we selected samples from patients demonstrating BJP by IEP in both serum and urine (n = 40), serum only (n = 18), and urine only (n = 32); 21 of these patients had BJP alone and 69 had BJP in addition to intact monoclonal Ig. Efficiency of detection of BJP in serum was increased by the present method: Serum BJP was detected in 70 patients by the present method versus 58 by IEP. The present method demonstrated single BJP bands in the samples from 16 patients (κ, n = 7; λ, n = 9) and multiple BJP bands (range: 2–9) in the samples from 54 patients (κ, n = 31; λ, n = 23). This method could be useful for detecting BJP in serum from patients suspected of having light chain gammopathy (without the need for urine testing) and may complement urine testing in patients excreting polyclonal free light chains of Ig. |
| doi_str_mv | 10.1177/000456329603300206 |
| format | Article |
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Treatment with PEG effectively reduced background staining of polyclonal Ig in the immunoblots, although intact monoclonal Ig was not always completely removed. To compare the present method with immunoelectrophoresis (IEP), we selected samples from patients demonstrating BJP by IEP in both serum and urine (n = 40), serum only (n = 18), and urine only (n = 32); 21 of these patients had BJP alone and 69 had BJP in addition to intact monoclonal Ig. Efficiency of detection of BJP in serum was increased by the present method: Serum BJP was detected in 70 patients by the present method versus 58 by IEP. The present method demonstrated single BJP bands in the samples from 16 patients (κ, n = 7; λ, n = 9) and multiple BJP bands (range: 2–9) in the samples from 54 patients (κ, n = 31; λ, n = 23). This method could be useful for detecting BJP in serum from patients suspected of having light chain gammopathy (without the need for urine testing) and may complement urine testing in patients excreting polyclonal free light chains of Ig.</description><identifier>ISSN: 0004-5632</identifier><identifier>EISSN: 1758-1001</identifier><identifier>DOI: 10.1177/000456329603300206</identifier><identifier>PMID: 8729721</identifier><identifier>CODEN: ACBOBU</identifier><language>eng</language><publisher>London, England: SAGE Publications</publisher><subject>Antibodies, Monoclonal ; Bence Jones Protein - analysis ; Bence Jones Protein - urine ; Biological and medical sciences ; Humans ; Immunoblotting ; Immunoelectrophoresis ; Immunoglobulins - isolation & purification ; Investigative techniques, diagnostic techniques (general aspects) ; Medical sciences ; Miscellaneous. Technology ; Paraproteinemias - metabolism ; Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques ; Polyethylene Glycols ; Reproducibility of Results ; Sensitivity and Specificity</subject><ispartof>Annals of clinical biochemistry, 1996-03, Vol.33 (2), p.132-138</ispartof><rights>1996 Association for Clinical Biochemistry</rights><rights>1996 INIST-CNRS</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c367t-a1e1c37edb19dd3c6183f8916ef08231c17418537c4ae113cf1059cb14d7f18e3</citedby><cites>FETCH-LOGICAL-c367t-a1e1c37edb19dd3c6183f8916ef08231c17418537c4ae113cf1059cb14d7f18e3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27901,27902</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&idt=3056103$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/8729721$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Backer, E T</creatorcontrib><creatorcontrib>Brand, A</creatorcontrib><title>Detection of Bence-Jones Protein in Serum by Immunoblotting</title><title>Annals of clinical biochemistry</title><addtitle>Ann Clin Biochem</addtitle><description>To detect Bence-Jones protein (BJP) in serum we precipitated intact immunoglobulins (Ig) using polyethylene glycol (PEG) and subjected the BJP in solution to electrophoresis in agarose gel, followed by transfer to a polyvinylidene difluoride membrane, and immunoenzymatic staining (successively using rabbit anti-human light/heavy chain of Ig, biotinylated swine anti-rabbit Ig, and alkaline phosphatase-conjugated streptavidin). Treatment with PEG effectively reduced background staining of polyclonal Ig in the immunoblots, although intact monoclonal Ig was not always completely removed. To compare the present method with immunoelectrophoresis (IEP), we selected samples from patients demonstrating BJP by IEP in both serum and urine (n = 40), serum only (n = 18), and urine only (n = 32); 21 of these patients had BJP alone and 69 had BJP in addition to intact monoclonal Ig. Efficiency of detection of BJP in serum was increased by the present method: Serum BJP was detected in 70 patients by the present method versus 58 by IEP. The present method demonstrated single BJP bands in the samples from 16 patients (κ, n = 7; λ, n = 9) and multiple BJP bands (range: 2–9) in the samples from 54 patients (κ, n = 31; λ, n = 23). This method could be useful for detecting BJP in serum from patients suspected of having light chain gammopathy (without the need for urine testing) and may complement urine testing in patients excreting polyclonal free light chains of Ig.</description><subject>Antibodies, Monoclonal</subject><subject>Bence Jones Protein - analysis</subject><subject>Bence Jones Protein - urine</subject><subject>Biological and medical sciences</subject><subject>Humans</subject><subject>Immunoblotting</subject><subject>Immunoelectrophoresis</subject><subject>Immunoglobulins - isolation & purification</subject><subject>Investigative techniques, diagnostic techniques (general aspects)</subject><subject>Medical sciences</subject><subject>Miscellaneous. Technology</subject><subject>Paraproteinemias - metabolism</subject><subject>Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques</subject><subject>Polyethylene Glycols</subject><subject>Reproducibility of Results</subject><subject>Sensitivity and Specificity</subject><issn>0004-5632</issn><issn>1758-1001</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>1996</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kE9Lw0AQxRdRaq1-AUHIQbzFzmST7AZPWv9VCgrqOWw2k5KS7Nbd5NBvb0pLL4IwMAzv997AY-wS4RZRiCkAxEnKoywFzgEiSI_YGEUiQwTAYzbeAuGWOGVn3q-GMxIAIzaSIspEhGN290gd6a62JrBV8EBGU_hmDfngw9mOahMM80mub4NiE8zbtje2aGzX1WZ5zk4q1Xi62O8J-35--pq9hov3l_nsfhFqnoouVEiouaCywKwsuU5R8kpmmFIFMuKoUcQoEy50rAiR6wohyXSBcSkqlMQn7GaXu3b2pyff5W3tNTWNMmR7nwuJPJMZDGC0A7Wz3juq8rWrW-U2OUK-bSz_29hgutqn90VL5cGyr2jQr_e68lo1lVNG1_6AcUhSHMImbLrDvFpSvrK9M0Mn_z3-BbfJfh8</recordid><startdate>19960301</startdate><enddate>19960301</enddate><creator>Backer, E T</creator><creator>Brand, A</creator><general>SAGE Publications</general><general>Royal Society of Medicine Press</general><scope>IQODW</scope><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>7X8</scope></search><sort><creationdate>19960301</creationdate><title>Detection of Bence-Jones Protein in Serum by Immunoblotting</title><author>Backer, E T ; Brand, A</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c367t-a1e1c37edb19dd3c6183f8916ef08231c17418537c4ae113cf1059cb14d7f18e3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>1996</creationdate><topic>Antibodies, Monoclonal</topic><topic>Bence Jones Protein - analysis</topic><topic>Bence Jones Protein - urine</topic><topic>Biological and medical sciences</topic><topic>Humans</topic><topic>Immunoblotting</topic><topic>Immunoelectrophoresis</topic><topic>Immunoglobulins - isolation & purification</topic><topic>Investigative techniques, diagnostic techniques (general aspects)</topic><topic>Medical sciences</topic><topic>Miscellaneous. Technology</topic><topic>Paraproteinemias - metabolism</topic><topic>Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques</topic><topic>Polyethylene Glycols</topic><topic>Reproducibility of Results</topic><topic>Sensitivity and Specificity</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Backer, E T</creatorcontrib><creatorcontrib>Brand, A</creatorcontrib><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Annals of clinical biochemistry</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Backer, E T</au><au>Brand, A</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Detection of Bence-Jones Protein in Serum by Immunoblotting</atitle><jtitle>Annals of clinical biochemistry</jtitle><addtitle>Ann Clin Biochem</addtitle><date>1996-03-01</date><risdate>1996</risdate><volume>33</volume><issue>2</issue><spage>132</spage><epage>138</epage><pages>132-138</pages><issn>0004-5632</issn><eissn>1758-1001</eissn><coden>ACBOBU</coden><abstract>To detect Bence-Jones protein (BJP) in serum we precipitated intact immunoglobulins (Ig) using polyethylene glycol (PEG) and subjected the BJP in solution to electrophoresis in agarose gel, followed by transfer to a polyvinylidene difluoride membrane, and immunoenzymatic staining (successively using rabbit anti-human light/heavy chain of Ig, biotinylated swine anti-rabbit Ig, and alkaline phosphatase-conjugated streptavidin). Treatment with PEG effectively reduced background staining of polyclonal Ig in the immunoblots, although intact monoclonal Ig was not always completely removed. To compare the present method with immunoelectrophoresis (IEP), we selected samples from patients demonstrating BJP by IEP in both serum and urine (n = 40), serum only (n = 18), and urine only (n = 32); 21 of these patients had BJP alone and 69 had BJP in addition to intact monoclonal Ig. Efficiency of detection of BJP in serum was increased by the present method: Serum BJP was detected in 70 patients by the present method versus 58 by IEP. The present method demonstrated single BJP bands in the samples from 16 patients (κ, n = 7; λ, n = 9) and multiple BJP bands (range: 2–9) in the samples from 54 patients (κ, n = 31; λ, n = 23). This method could be useful for detecting BJP in serum from patients suspected of having light chain gammopathy (without the need for urine testing) and may complement urine testing in patients excreting polyclonal free light chains of Ig.</abstract><cop>London, England</cop><pub>SAGE Publications</pub><pmid>8729721</pmid><doi>10.1177/000456329603300206</doi><tpages>7</tpages></addata></record> |
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| ispartof | Annals of clinical biochemistry, 1996-03, Vol.33 (2), p.132-138 |
| issn | 0004-5632 1758-1001 |
| language | eng |
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| source | MEDLINE; Alma/SFX Local Collection |
| subjects | Antibodies, Monoclonal Bence Jones Protein - analysis Bence Jones Protein - urine Biological and medical sciences Humans Immunoblotting Immunoelectrophoresis Immunoglobulins - isolation & purification Investigative techniques, diagnostic techniques (general aspects) Medical sciences Miscellaneous. Technology Paraproteinemias - metabolism Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques Polyethylene Glycols Reproducibility of Results Sensitivity and Specificity |
| title | Detection of Bence-Jones Protein in Serum by Immunoblotting |
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