Source:http://linkedlifedata.com/resource/pubmed/id/18242632
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| Predicate | Object |
|---|---|
| rdf:type | |
| lifeskim:mentions | |
| pubmed:issue |
1-2
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| pubmed:dateCreated |
2008-3-17
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| pubmed:abstractText |
Methylglyoxal (MG) is generated through the Embden-Meyerhof and polyol pathways, and it rapidly reacts with proteins to form advanced glycation end products (AGE) such as N(epsilon)-(carboxyethyl)lysine (CEL). In the present study, polyclonal and monoclonal antibodies specific for CEL were prepared to estimate CEL content in aldehydes-modified proteins and the pathological localization in human kidneys. Polyclonal CEL-specific antibody was prepared by removing cross-reactive antibodies against N(epsilon)-(carboxymethyl) lysine (CML), one of the major AGE structures, using CML-conjugated affinity chromatography. Monoclonal CEL-specific antibody (CEL-SP) was obtained by immunization with CEL-bovine serum albumin, followed by successive screening according to CEL-RNase-positive but CML-RNase-negative criteria. A non-competitive ELISA showed that both the polyclonal and monoclonal CEL-specific antibodies significantly reacted with CEL-proteins but not with CML-proteins. A competitive ELISA also demonstrated that CEL-SP does not show cross-reactivity against CEL analogues such as CML, carboxymethylarginine (CMA) and S-carboxymethylcysteine (CMC), thus indicating that antibody is able to recognize the difference of one methyl group between carboxymethyl group and carboxyethyl group. Furthermore, CEL-SP significantly reacted with human serum albumin modified with MG but not with glyoxal or 3-deoxyglucosone, and its reactivity was highly correlated with the CEL content, which was determined by high performance liquid chromatography. Immunohistochemical studies using CEL-SP provided evidence that CEL-modified proteins accumulate in distal tubular epithelial cells of the diabetic rat. These results demonstrate that a specific antibody against CEL can be a powerful tool for detecting CEL both in vitro and in vivo.
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| pubmed:language |
eng
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| pubmed:journal | |
| pubmed:citationSubset |
IM
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| pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/Aldehydes,
http://linkedlifedata.com/resource/pubmed/chemical/Antibodies, Monoclonal,
http://linkedlifedata.com/resource/pubmed/chemical/Lysine,
http://linkedlifedata.com/resource/pubmed/chemical/N(6)-carboxymethyllysine,
http://linkedlifedata.com/resource/pubmed/chemical/Pyruvaldehyde,
http://linkedlifedata.com/resource/pubmed/chemical/Serum Albumin
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| pubmed:status |
MEDLINE
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| pubmed:month |
Mar
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| pubmed:issn |
0022-1759
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| pubmed:author | |
| pubmed:issnType |
Print
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| pubmed:day |
20
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| pubmed:volume |
332
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| pubmed:owner |
NLM
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| pubmed:authorsComplete |
Y
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| pubmed:pagination |
112-20
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| pubmed:meshHeading |
pubmed-meshheading:18242632-Aldehydes,
pubmed-meshheading:18242632-Animals,
pubmed-meshheading:18242632-Antibodies, Monoclonal,
pubmed-meshheading:18242632-Antibody Specificity,
pubmed-meshheading:18242632-Antigen-Antibody Reactions,
pubmed-meshheading:18242632-Cattle,
pubmed-meshheading:18242632-Chromatography, High Pressure Liquid,
pubmed-meshheading:18242632-Diabetes Mellitus, Experimental,
pubmed-meshheading:18242632-Enzyme-Linked Immunosorbent Assay,
pubmed-meshheading:18242632-Humans,
pubmed-meshheading:18242632-Immunohistochemistry,
pubmed-meshheading:18242632-Kidney,
pubmed-meshheading:18242632-Lysine,
pubmed-meshheading:18242632-Male,
pubmed-meshheading:18242632-Pyruvaldehyde,
pubmed-meshheading:18242632-Rats,
pubmed-meshheading:18242632-Rats, Wistar,
pubmed-meshheading:18242632-Sensitivity and Specificity,
pubmed-meshheading:18242632-Serum Albumin
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| pubmed:year |
2008
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| pubmed:articleTitle |
Immunochemical detection of Nepsilon-(carboxyethyl)lysine using a specific antibody.
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| pubmed:affiliation |
Department of Medical Biochemistry, Faculty of Medical and Pharmaceutical Sciences, Kumamoto University, Kumamoto, Japan. nagai-883@umin.ac.jp
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| pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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