{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,5]],"date-time":"2026-05-05T00:46:18Z","timestamp":1777941978380,"version":"3.51.4"},"reference-count":21,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2012,1,30]],"date-time":"2012-01-30T00:00:00Z","timestamp":1327881600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["J of Molecular Recognition"],"published-print":{"date-parts":[[2012,2]]},"abstract":"<jats:p>Antibodies are a promising tool for the fast and selective trace detection of explosives. Unfortunately, the production of high\u2010quality antibodies is not trivial and often expensive. Therefore, excellent antibodies are a rare and limiting resource in fields such as biosensing, environmental analysis, diagnostics, cancer therapy, and proteomics. Here, we report the synthesis, bioconjugation, and application of the structurally optimized hapten 6\u2010(2,4,6\u2010trinitro)\u2010phenylhexanoic acid to improve the selectivity and sensitivity of antibodies for the detection of one of the most important explosives, trinitrotoluene. With a conjugate of bovine serum albumin and a highly purified <jats:italic>N<\/jats:italic>\u2010hydroxy\u2010succinimide (NHS)\u2013activated hapten, two rabbits were immunized to obtain polyclonal antibodies. The immunization process was monitored by enzyme\u2010linked immunosorbent assay to gain information about the progress of antibody titer and affinity. Finally, the polyclonal antibodies reached an affinity constant of (5.1\u2009\u00b1\u20090.6)\u2009\u00d7\u200910<jats:sup>9<\/jats:sup>\u2009l\/mol (rabbit R1) and (2.3\u2009\u00b1\u20090.2)\u2009\u00d7\u200910<jats:sup>9<\/jats:sup>\u2009l\/mol (rabbit R2). The respective assays show a minimum test midpoint (IC<jats:sub>50<\/jats:sub> value) of 0.1\u2009\u00b1\u20090.01\u2009\u00b5g\/l (R1) and 0.2\u2009\u00b1\u20090.02\u2009\u00b5g\/l (R2) and a working range of 0.005 to 150\u2009\u00b5g\/l (R1) and 0.007 to 200\u2009\u00b5g\/l (R2), which corresponds to more than four orders of magnitude for both. This is quite remarkable for a competitive immunoassay, which is often believed to have a narrow dynamic range. The limit of detection was calculated to 0.6\u2009ng\/l (R1) and 1.5\u2009ng\/l (R2), which is up to 100 times improvement in relation to the assay of Zeck <jats:italic>et al.<\/jats:italic> (1999) on the basis of a monoclonal antibody. The excellent selectivity of the polyclonal antibodies was comprehensively examined by determining the cross\u2010reactivity to common explosives and other nitroaromatics including nitro musk components. The widely held belief that polyclonal antibodies generally display higher cross\u2010reactivities than monoclonals could be disproved. Copyright \u00a9 2012 John Wiley &amp; Sons, Ltd.<\/jats:p>","DOI":"10.1002\/jmr.2162","type":"journal-article","created":{"date-parts":[[2012,1,30]],"date-time":"2012-01-30T18:56:27Z","timestamp":1327949787000},"page":"89-97","source":"Crossref","is-referenced-by-count":28,"title":["Extremely sensitive and selective antibodies against the explosive 2,4,6\u2010trinitrotoluene by rational design of a structurally optimized hapten"],"prefix":"10.1002","volume":"25","author":[{"given":"Steffen","family":"Ramin","sequence":"first","affiliation":[{"name":"BAM Federal Institute for Materials Research and Testing Division 1.5, Protein Analysis  Richard\u2010Willst\u00e4tter\u2010Strasse 11 12489 Berlin Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael G.","family":"Weller","sequence":"additional","affiliation":[{"name":"BAM Federal Institute for Materials Research and Testing Division 1.5, Protein Analysis  Richard\u2010Willst\u00e4tter\u2010Strasse 11 12489 Berlin Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2012,1,30]]},"reference":[{"key":"e_1_2_7_2_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00216-009-2958-7"},{"key":"e_1_2_7_3_1","doi-asserted-by":"publisher","DOI":"10.1016\/0022-4731(74)90061-2"},{"key":"e_1_2_7_4_1","doi-asserted-by":"publisher","DOI":"10.1126\/science.320.5882.1416"},{"key":"e_1_2_7_5_1","doi-asserted-by":"publisher","DOI":"10.1023\/A:1015890208714"},{"key":"e_1_2_7_6_1","doi-asserted-by":"publisher","DOI":"10.1039\/b200627h"},{"key":"e_1_2_7_7_1","first-page":"33","article-title":"The \u201cPrecision Profile\u201d: Its use in RIA assessment and design","volume":"4","author":"Ekins R","year":"1981","journal-title":"The Ligand Quarterly"},{"key":"e_1_2_7_8_1","doi-asserted-by":"publisher","DOI":"10.1021\/es034328e"},{"key":"e_1_2_7_9_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00216-005-3294-1"},{"key":"e_1_2_7_10_1","doi-asserted-by":"publisher","DOI":"10.1021\/bi00875a001"},{"key":"e_1_2_7_11_1","doi-asserted-by":"publisher","DOI":"10.1016\/0165-9936(95)90920-I"},{"key":"e_1_2_7_12_1","doi-asserted-by":"crossref","first-page":"381","DOI":"10.1093\/jaoac\/76.2.381","article-title":"Development of enzyme\u2010immunoassay for captan and its degradation product tetrahydrophthalimide in foods","volume":"76","author":"Newsome WH","year":"1993","journal-title":"J. AOAC Int."},{"key":"e_1_2_7_13_1","doi-asserted-by":"publisher","DOI":"10.1016\/0003-2697(78)90002-7"},{"key":"e_1_2_7_14_1","doi-asserted-by":"publisher","DOI":"10.1039\/b717933m"},{"key":"e_1_2_7_15_1","first-page":"38","volume-title":"New frontiers in agrochemical immunoassay","author":"Szurdoki F","year":"1995"},{"key":"e_1_2_7_16_1","doi-asserted-by":"publisher","DOI":"10.3390\/bios1030093"},{"key":"e_1_2_7_17_1","unstructured":"WellerMG.1992.Strukturelle und kinetische Untersuchungen zur Entwicklung und Optimierung von Hapten\u2010Enzymimmunoassays (ELISAs) am Beispiel der Bestimmung von Triazinherbiziden. PhD Thesis. Technische Universit\u00e4t M\u00fcnchen (German)."},{"key":"e_1_2_7_18_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0003-2670(98)00855-1"},{"key":"e_1_2_7_19_1","doi-asserted-by":"publisher","DOI":"10.1002\/jlac.18631280206"},{"key":"e_1_2_7_20_1","doi-asserted-by":"publisher","DOI":"10.1007\/s002160051309"},{"key":"e_1_2_7_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0003-2670(01)01092-3"},{"key":"e_1_2_7_22_1","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1093\/clinchem\/20.1.5","article-title":"Principles of competitive binding assays (saturation analyses). 2. Sequential saturation","volume":"20","author":"Zettner A","year":"1974","journal-title":"Clin. Chem."}],"container-title":["Journal of Molecular Recognition"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1002%2Fjmr.2162","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1002\/jmr.2162","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,9,23]],"date-time":"2023-09-23T18:08:11Z","timestamp":1695492491000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1002\/jmr.2162"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2012,1,30]]},"references-count":21,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2012,2]]}},"alternative-id":["10.1002\/jmr.2162"],"URL":"https:\/\/doi.org\/10.1002\/jmr.2162","archive":["Portico"],"relation":{},"ISSN":["0952-3499","1099-1352"],"issn-type":[{"value":"0952-3499","type":"print"},{"value":"1099-1352","type":"electronic"}],"subject":[],"published":{"date-parts":[[2012,1,30]]}}}