{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2024,7,31]],"date-time":"2024-07-31T22:32:58Z","timestamp":1722465178137},"reference-count":21,"publisher":"Cambridge University Press (CUP)","issue":"1","license":[{"start":{"date-parts":[[2014,12,22]],"date-time":"2014-12-22T00:00:00Z","timestamp":1419206400000},"content-version":"unspecified","delay-in-days":0,"URL":"https:\/\/www.cambridge.org\/core\/terms"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["J. Helminthol."],"published-print":{"date-parts":[[2016,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Global programmes to eliminate lymphatic filariasis (GPELF) require mapping, monitoring and evaluation using filarial antigen diagnostic kits. To meet this objective, a functional single-chain fragment variable (ScFv) specific for filarial <jats:italic>Wuchereria bancrofti<\/jats:italic> SXP-1 (<jats:italic>Wb<\/jats:italic>-SXP-1) antigen was constructed for the diagnosis of active filarial infection, an alternative to the production of complete antibodies using hybridomas. The variable heavy chain (V<jats:sub>H<\/jats:sub>) and the variable light chain (kappa) (V\u03ba) genes were amplified from the mouse hybridoma cell line and were linked together with a flexible linker by overlap extension polymerase chain reaction (PCR). The ScFv construct (V\u03ba\u2013Linker\u2013V<jats:sub>H<\/jats:sub>) was expressed as a fusion protein with N-terminal His tag in <jats:italic>Escherichia coli<\/jats:italic> and purified using immobilized metal affinity chromatography (IMAC) without the addition of reducing agents. Immunoblotting and sandwich enzyme-linked immunosorbent assay (ELISA) were used to analyse the antigen binding affinity of purified ScFv. The purified ScFv was found to recognize recombinant and native <jats:italic>Wb<\/jats:italic>-SXP-1 antigen in microfilariae (Mf)-positive patient sera. The affinity of ScFv was comparable with that of the monoclonal antibody. The development of recombinant ScFv to replace monoclonal antibody for detection of filarial antigen was achieved. The recombinant ScFv was purified, on-column refolded and its detection ability validated using field samples.<\/jats:p>","DOI":"10.1017\/s0022149x14000868","type":"journal-article","created":{"date-parts":[[2014,12,22]],"date-time":"2014-12-22T06:28:12Z","timestamp":1419229692000},"page":"74-80","source":"Crossref","is-referenced-by-count":1,"title":["Construction and bacterial expression of a recombinant single-chain antibody fragment against <i>Wuchereria bancrofti<\/i> SXP-1 antigen for the diagnosis of lymphatic filariasis"],"prefix":"10.1017","volume":"90","author":[{"given":"R.","family":"Kamatchi","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"J.","family":"Charumathi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"R.","family":"Ravishankaran","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"P.","family":"Kaliraj","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"S.","family":"Meenakshisundaram","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"56","published-online":{"date-parts":[[2014,12,22]]},"reference":[{"key":"S0022149X14000868_ref20","doi-asserted-by":"publisher","DOI":"10.4269\/ajtmh.13-0089"},{"key":"S0022149X14000868_ref19","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-1759(99)00184-2"},{"key":"S0022149X14000868_ref18","doi-asserted-by":"publisher","DOI":"10.1017\/S0022149X00015157"},{"key":"S0022149X14000868_ref14","doi-asserted-by":"publisher","DOI":"10.1016\/j.cca.2011.06.029"},{"key":"S0022149X14000868_ref13","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.86.10.3833"},{"key":"S0022149X14000868_ref12","first-page":"253","article-title":"Comparative assessment of an Og4C3 ELISA and an ICT filariasis test: A study of Myanmar migrants in Thailand","volume":"21","author":"Nuchprayoon","year":"2003","journal-title":"Asian Pacific Journal of Allergy and Immunology"},{"key":"S0022149X14000868_ref9","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-3156.1998.00170.x"},{"key":"S0022149X14000868_ref8","volume-title":"Sequences of proteins of immunological interest","author":"Kabat","year":"1987"},{"key":"S0022149X14000868_ref4","doi-asserted-by":"publisher","DOI":"10.1038\/nbt1142"},{"key":"S0022149X14000868_ref3","doi-asserted-by":"publisher","DOI":"10.1021\/bi00458a002"},{"key":"S0022149X14000868_ref2","doi-asserted-by":"publisher","DOI":"10.1007\/BF00176675"},{"key":"S0022149X14000868_ref17","doi-asserted-by":"publisher","DOI":"10.1016\/j.copbio.2006.05.011"},{"key":"S0022149X14000868_ref10","doi-asserted-by":"publisher","DOI":"10.1111\/j.1348-0421.2002.tb02703.x"},{"key":"S0022149X14000868_ref21","doi-asserted-by":"publisher","DOI":"10.1007\/s00253-009-2379-8"},{"key":"S0022149X14000868_ref7","doi-asserted-by":"publisher","DOI":"10.1017\/S0022149X10000118"},{"key":"S0022149X14000868_ref5","doi-asserted-by":"publisher","DOI":"10.1016\/S0958-1669(98)80014-1"},{"key":"S0022149X14000868_ref6","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.85.16.5879"},{"key":"S0022149X14000868_ref16","volume-title":"Molecular cloning: a laboratory manual","author":"Sambrook","year":"2001"},{"key":"S0022149X14000868_ref15","doi-asserted-by":"publisher","DOI":"10.1016\/S0166-6851(99)00231-5"},{"key":"S0022149X14000868_ref11","doi-asserted-by":"publisher","DOI":"10.1006\/prep.2002.1623"},{"key":"S0022149X14000868_ref1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.71.3.845"}],"container-title":["Journal of Helminthology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.cambridge.org\/core\/services\/aop-cambridge-core\/content\/view\/S0022149X14000868","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2019,4,19]],"date-time":"2019-04-19T19:48:31Z","timestamp":1555703311000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.cambridge.org\/core\/product\/identifier\/S0022149X14000868\/type\/journal_article"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2014,12,22]]},"references-count":21,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2016,1]]}},"alternative-id":["S0022149X14000868"],"URL":"https:\/\/doi.org\/10.1017\/s0022149x14000868","relation":{},"ISSN":["0022-149X","1475-2697"],"issn-type":[{"value":"0022-149X","type":"print"},{"value":"1475-2697","type":"electronic"}],"subject":[],"published":{"date-parts":[[2014,12,22]]}}}