{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,2,3]],"date-time":"2026-02-03T02:24:00Z","timestamp":1770085440835,"version":"3.49.0"},"reference-count":32,"publisher":"Oxford University Press (OUP)","issue":"2","license":[{"start":{"date-parts":[[2001,1,1]],"date-time":"2001-01-01T00:00:00Z","timestamp":978307200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2001,1,15]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>We describe a two-stage preparation of chemically engineered Ab constructs, employing as modules Fab\u2032\u03b3 from mAb or rAb, and Fc from human normal IgG1. A multivalent, optionally multispecific F(ab\u2032)n core is formed in stage one, and one or more Fc modules added in stage two. Examples include bispecific Fab2Fc2 (for simplicity, primes and Greek letters are omitted from names of final constructs) and trivalent Fab3Fc2, which are designed to kill neoplastic cells. An essential element in the construction is the availability of the Fab\u2032 in two reduced forms, Fab\u2032(-sulfhydryl (SH))5 and Fab\u2032-SH. The first is obtained by full reduction of the interchain disulfide bonds (SS) in the F(ab\u2032)2 fragment of IgG. Fab\u2032-SH is obtained by disulfide-interchange reactions on Fab\u2032(-SH)5, whereby the \u03b3-light SS is reconstituted, an unusual intrachain SS forms in the \u03b3-chain hinge, and one hinge SH remains. F(ab\u2032)2 and F(ab\u2032)3 cores are built using partially reduced modules, being given intermodular thioether links that resist reduction. These cores are then fully reduced, making available SH groups for addition of the Fc\u03b3 modules. In the final constructs, all intermodular links embody tandem thioether bonds arising at hinge-region cysteines. Cytotoxic activities of representative constructs, and some enhancements deriving from multiple modules, are assessed. In guinea pigs, catabolism of Fab2Fc2 yielded a t1\/2 similar to that of human IgG1, although the serum Fab2Fc2 revealed some proteolytic breakdown not shown by the IgG1. Immunotherapy of a guinea-pig leukemia confirmed the ability of these constructs to kill target cells in vivo.<\/jats:p>","DOI":"10.4049\/jimmunol.166.2.1320","type":"journal-article","created":{"date-parts":[[2014,4,20]],"date-time":"2014-04-20T22:56:47Z","timestamp":1398034607000},"page":"1320-1326","source":"Crossref","is-referenced-by-count":6,"title":["Thioether-Bonded Constructs of Fab\u2032\u03b3 and Fc\u03b3 Modules Utilizing Differential Reduction of Interchain Disulfide Bonds"],"prefix":"10.1093","volume":"166","author":[{"given":"Kwok S","family":"Kan","sequence":"first","affiliation":[{"name":"Biological Sciences, University of North London , London ,","place":["United Kingdom"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Valerie A","family":"Anderson","sequence":"additional","affiliation":[{"name":"Tenovus Research Laboratory, Southampton University Hospitals , Southampton ,","place":["United Kingdom"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Weng S","family":"Leong","sequence":"additional","affiliation":[{"name":"Tenovus Research Laboratory, Southampton University Hospitals , Southampton ,","place":["United Kingdom"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andrew M","family":"Smith","sequence":"additional","affiliation":[{"name":"Tenovus Research Laboratory, Southampton University Hospitals , Southampton ,","place":["United Kingdom"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andrew T","family":"Worth","sequence":"additional","affiliation":[{"name":"The Edward Jenner Institute for Vaccine , Compton ,","place":["United Kingdom"]}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"George T","family":"Stevenson","sequence":"additional","affiliation":[{"name":"Tenovus Research Laboratory, Southampton University Hospitals , Southampton ,","place":["United Kingdom"]}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"286","published-online":{"date-parts":[[2001,1,15]]},"reference":[{"key":"2026020208094393000_R1"},{"key":"2026020208094393000_R2","first-page":"2367","volume-title":"J. 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