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publicationDate 1993-09-02-04:00^^<http://www.w3.org/2001/XMLSchema#date>
publicationNumber DE-4205938-A1
titleOfInvention IMMUNOTOXINE
abstract New immunotoxin (I) comprises monoclonal CD30 antibodies (Ab) covalently bonded to a ribosome-inactivating protein type-1 toxin (II). Pref. Ab is of the Ber-H2 type while (II) is bryodin; the toxin purified from Asparagus officinalis; momorcochin-5; dianthin; gelonin; trichosanthin or pref. saporin; PAP (from Phytolacca americana seeds) and/or momordin. Ab, esp. Ber-H2 from the hybridoma ECACC 92012823, and (II) are coupled using a heterobifunctional reagent, e.g. N-succininidyl-3-(2-pyridyldithio)propionate; S-acetyl-mercaptosuccinic anhydride; a carbodiimide; glutaraldehyde or esp. 2-iminothiolan (2-IT; sec Br. J cancer, 60 (1989) 315). Ab is isolated from hybridoma culture supernatant (or ascites) by affinity chromatography on protein A and then hydroxyapatite chromatography. It is derivatised with 2-IT in pH9 borate buffer then incubated for 20 hr at room temp. with 2-IT-derivatised toxin (reduced with dithiothreitol). The reaction mixt. was gel. filtered on 'Sephacryl S-200' (RTM) and the fractions contg. the low molecular wt. conjugate isolated (esp. it has mole ratio (II): Ab = 1-3). USE/ADVANTAGE - (I) can be used (1) for treating tumours and (2) for selective elimination, in vivo or ex vivo, of CD-30 positive cells. They are more active than similar immuno toxins prepd. from ricin A chain and give partic. good results in cases of Hodgkin's disease. (I) is administered by intramuscular or intravenous injection (as a soln. or suspension) to provide a dose of 0.01-5 (pref. 0.01-0.5) mg/kg, given one or more times per day for one to several weeks.
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