{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T15:48:45Z","timestamp":1792165725368,"version":"4.4.0"},"reference-count":17,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2006,7,27]],"date-time":"2006-07-27T00:00:00Z","timestamp":1153958400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["European J of Haematology"],"published-print":{"date-parts":[[2006,11]]},"abstract":"<jats:p><jats:bold>Abstract:\u2002<\/jats:bold>  <jats:italic>Objectives:<\/jats:italic>\u2002Individual immunoglobulins expressed by B\u2010cell lymphomas represent tumor\u2010specific antigens (\u2018idiotypes\u2019). Immunization with idiotype in follicular lymphoma patients may induce specific immune responses, sustained progression\u2010free survival, and disappearance of minimal residual disease. Manufacturing of idiotype vaccines has mostly relied on heterohybridomas established from viable lymphoma cells. This paper describes the feasibility of production of GMP\u2010grade idiotype vaccines as recombinant Fab fragments in <jats:italic>Escherichia coli<\/jats:italic>.<\/jats:p><jats:p> <jats:italic>Methods:<\/jats:italic>\u2002IgH and IgL transcripts were analyzed by anchored PCR from 106 lymphoma and nine control biopsies. Lymphoma\u2010derived V segments were inserted into prokaryotic expression plasmids. Recombinant idiotype Fab fragments were expressed in <jats:italic>E. coli<\/jats:italic> in a fermentation system.<\/jats:p><jats:p> <jats:italic>Results:<\/jats:italic>\u2002Idiotype IgH and IgL transcripts were identified in 95% of 106 lymphoma biopsies according to stringent clonality criteria. Large\u2010scale idiotype expression was successful in 69 of 78 cases (89%) and yielded a median of 17\u2003mg (range: 1.2\u2013250\u2003mg) recombinant Fab protein. After affinity chromatography, median vaccine purity was 99% heterodimeric Fab protein (range: 72\u2013100%). Bacterial protein contamination was detectable in one vaccine only. Fab proteins with IgL lambda chains had a tendency for inferior yield and lesser purity than <jats:italic>\u03ba<\/jats:italic>\u2010type Fabs. Among other structural idiotype features (isotype, V family usage, somatic hypermutation pattern, novel glycosylation sites, CDR III net charge), no consistent influences on Fab yield or purity were detected.<\/jats:p><jats:p> <jats:italic>Conclusions:<\/jats:italic>\u2002Anchored PCR cloning and subsequent protein expression in <jats:italic>E. coli<\/jats:italic> provides a reliable technological basis for clinical idiotype vaccination trials.<\/jats:p>","DOI":"10.1111\/j.1600-0609.2006.00740.x","type":"journal-article","created":{"date-parts":[[2006,7,27]],"date-time":"2006-07-27T11:00:40Z","timestamp":1153998040000},"page":"395-402","source":"Crossref","is-referenced-by-count":21,"title":["Cloning of idiotype immunoglobulin genes in B cell lymphomas by anchored PCR and production of individual recombinant idiotype vaccines in <i>Escherichia coli<\/i>"],"prefix":"10.1111","volume":"77","author":[{"given":"Cristina","family":"Bertinetti","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Frederic","family":"Simon","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katja","family":"Zirlik","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kristina","family":"Heining\u2010Mikesch","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dietmar","family":"Pfeifer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Frank","family":"Osterroth","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Felicia M.","family":"Rosenthal","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hendrik","family":"Veelken","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"311","published-online":{"date-parts":[[2006,7,27]]},"reference":[{"key":"e_1_2_6_2_2","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V95.4.1342.004k19_1342_1349"},{"key":"e_1_2_6_3_2","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V89.9.3129"},{"key":"e_1_2_6_4_2","doi-asserted-by":"publisher","DOI":"10.1038\/13928"},{"key":"e_1_2_6_5_2","doi-asserted-by":"publisher","DOI":"10.1046\/j.1365-2141.1996.419959.x"},{"key":"e_1_2_6_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/0022-1759(86)90032-3"},{"key":"e_1_2_6_7_2","doi-asserted-by":"crossref","first-page":"3279","DOI":"10.1182\/blood.V83.11.3279.3279","article-title":"Idiotypic vaccination against human B\u2010cell lymphoma. Rescue of variable region gene sequences from biopsy material for assembly as single\u2010chain Fv personal vaccines","volume":"83","author":"Hawkins RE","year":"1994","journal-title":"Blood"},{"key":"e_1_2_6_8_2","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V99.5.1517"},{"key":"e_1_2_6_9_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0022-1759(99)00111-8"},{"key":"e_1_2_6_10_2","doi-asserted-by":"publisher","DOI":"10.1158\/0008-5472.CAN-05-4233"},{"key":"e_1_2_6_11_2","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V99.7.2562"},{"key":"e_1_2_6_12_2","doi-asserted-by":"crossref","first-page":"2922","DOI":"10.1182\/blood.V95.9.2922.009k38_2922_2929","article-title":"Variable heavy chain gene analysis of follicular lymphomas: correlation between heavy chain isotype expression and somatic mutation load","volume":"95","author":"Aarts WM","year":"2000","journal-title":"Blood"},{"key":"e_1_2_6_13_2","doi-asserted-by":"publisher","DOI":"10.1038\/sj.leu.2403202"},{"key":"e_1_2_6_14_2","doi-asserted-by":"publisher","DOI":"10.1038\/sj.leu.2400801"},{"key":"e_1_2_6_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/0046-8177(94)90085-X"},{"key":"e_1_2_6_16_2","first-page":"1438","article-title":"Feasibility of idiotype vaccination in relapsed B\u2010cell malignancies","volume":"88","author":"Inoges S","year":"2003","journal-title":"Haematologica"},{"key":"e_1_2_6_17_2","doi-asserted-by":"publisher","DOI":"10.1084\/jem.188.4.773"},{"key":"e_1_2_6_18_2","doi-asserted-by":"publisher","DOI":"10.1084\/jem.186.4.479"}],"container-title":["European Journal of Haematology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1111%2Fj.1600-0609.2006.00740.x","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/j.1600-0609.2006.00740.x","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,16]],"date-time":"2023-10-16T03:49:13Z","timestamp":1697428153000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/j.1600-0609.2006.00740.x"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2006,7,27]]},"references-count":17,"journal-issue":{"issue":"5","published-print":{"date-parts":[[2006,11]]}},"alternative-id":["10.1111\/j.1600-0609.2006.00740.x"],"URL":"https:\/\/doi.org\/10.1111\/j.1600-0609.2006.00740.x","archive":["Portico"],"relation":{},"ISSN":["0902-4441","1600-0609"],"issn-type":[{"value":"0902-4441","type":"print"},{"value":"1600-0609","type":"electronic"}],"subject":[],"published":{"date-parts":[[2006,7,27]]}}}