{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,27]],"date-time":"2025-10-27T04:57:31Z","timestamp":1761541051632},"reference-count":0,"publisher":"Ovid Technologies (Wolters Kluwer Health)","issue":"6","content-domain":{"domain":["www.ahajournals.org"],"crossmark-restriction":true},"short-container-title":["Circulation"],"published-print":{"date-parts":[[1990,6]]},"abstract":"<jats:p>Single-chain urokinase plasminogen activator (scu-PA) that had been modified with N-succinimidyl-3-(2-pyridyldithio)propionate was covalently linked by disulfide bonds to the Fab' of a monoclonal antibody specific for the beta-chain of fibrin (antibody 59D8). scu-PA-59D8 Fab' conjugate was separated from free scu-PA and two-chain urokinase coupled to 59D8 Fab' by two-step affinity chromatography. First, the reaction mixture was chromatographed on a column containing Sepharose linked to the peptide that had been used as immunogen for antibody 59D8; scu-PA-59D8 Fab' conjugate and unconjugated 59D8 Fab' were retained but not unconjugated scu-PA. Then, the eluate from the peptide-Sepharose column was chromatographed on a column containing Sepharose linked to benzamidine, which acts as a ligand for two-chain urokinase. The molecular weight of the scu-PA-59D8 Fab' conjugate was approximately 100 kDa when electrophoresed on a nonreducing sodium dodecylsulfate-polyacrylamide gel. Enzymatic assay after purification revealed that more than 97% of the scu-PA present in the conjugate retained the single-chain form. The Fab' portion of the conjugate functioned in a manner indistinguishable from that of native antibody 59D8. In an in vitro assay for lysis of fibrin monomer, the fibrinolytic potency of scu-PA-59D8 Fab' was 33-fold more than that of tissue plasminogen activator (p less than 0.001), 230-fold more than that of unconjugated scu-PA (p less than 0.0001), and 420-fold more than that of urokinase (p less than 0.0001).(ABSTRACT TRUNCATED AT 250 WORDS)<\/jats:p>","DOI":"10.1161\/01.cir.81.6.1974","type":"journal-article","created":{"date-parts":[[2011,6,17]],"date-time":"2011-06-17T20:29:13Z","timestamp":1308342553000},"page":"1974-1980","update-policy":"http:\/\/dx.doi.org\/10.1161\/crossmarkpolicy","source":"Crossref","is-referenced-by-count":38,"title":["Conjugation to antifibrin Fab' enhances fibrinolytic potency of single-chain urokinase plasminogen activator."],"prefix":"10.1161","volume":"81","author":[{"given":"C","family":"Bode","sequence":"first","affiliation":[{"name":"Medizinische Klinik III (Kardiologie), Universit\u00e4t Heidelberg, FRG."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"M S","family":"Runge","sequence":"additional","affiliation":[{"name":"Medizinische Klinik III (Kardiologie), Universit\u00e4t Heidelberg, FRG."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"S","family":"Sch\u00f6nermark","sequence":"additional","affiliation":[{"name":"Medizinische Klinik III (Kardiologie), Universit\u00e4t Heidelberg, FRG."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"T","family":"Eberle","sequence":"additional","affiliation":[{"name":"Medizinische Klinik III (Kardiologie), Universit\u00e4t Heidelberg, FRG."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"J B","family":"Newell","sequence":"additional","affiliation":[{"name":"Medizinische Klinik III (Kardiologie), Universit\u00e4t Heidelberg, FRG."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"W","family":"K\u00fcbler","sequence":"additional","affiliation":[{"name":"Medizinische Klinik III (Kardiologie), Universit\u00e4t Heidelberg, FRG."}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"E","family":"Haber","sequence":"additional","affiliation":[{"name":"Medizinische Klinik III (Kardiologie), Universit\u00e4t Heidelberg, FRG."}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"276","container-title":["Circulation"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.ahajournals.org\/doi\/pdf\/10.1161\/01.CIR.81.6.1974","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,5,12]],"date-time":"2024-05-12T16:07:31Z","timestamp":1715530051000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.ahajournals.org\/doi\/10.1161\/01.CIR.81.6.1974"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[1990,6]]},"references-count":0,"journal-issue":{"issue":"6","published-print":{"date-parts":[[1990,6]]}},"alternative-id":["10.1161\/01.CIR.81.6.1974"],"URL":"https:\/\/doi.org\/10.1161\/01.cir.81.6.1974","relation":{},"ISSN":["0009-7322","1524-4539"],"issn-type":[{"value":"0009-7322","type":"print"},{"value":"1524-4539","type":"electronic"}],"subject":[],"published":{"date-parts":[[1990,6]]},"assertion":[{"value":"1990-06-01","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}]}}