MORAN JUSTIN KEITH,RUPPEN MARK EDWARD,POPPLEWELL ANDREW GEORGE,DIJOSEPH JOHN FRANCIS,DAMLE NITIN KRISHNAJI,SIMPSON JOHN MCLEAN,MERCHANT NISHITH,ROBBINS PAUL DAVID,RUBINO JOSEPH THOMAS,VIDUNAS EUGENE J
申请号：
NZ61195103
公开号：
NZ611951A
申请日:
2003.05.02
申请国别(地区):
NZ
年份:
2014
代理人:
摘要：
Disclosed is a method for preparing a composition with reduced low conjugated fraction (LCF) below 10% comprising monomeric calicheamicin derivative/antibody conjugates having the formula, Pr(-X-W)m wherein: Pr is an antibody X is a hydrolyzable linker having the formula (CO-Alk1-sp1-Ar-sp2-Alk2-C(Z1)=Q-Sp) as defined herein the method comprising the steps of: (1) adding the calicheamicin derivative to the antibody wherein the calicheamicin derivative is 4.5-11% by weight of the antibody (2) incubating the calicheamicin derivative and the antibody in a non-nucleophilic, protein-compatible, buffered solution having a pH in the range from about 7 to 9 to produce a monomeric calicheamicin derivative/antibody conjugate, wherein the solution further comprises (a) an organic cosolvent, and (b) an additive comprising 5 at least one C6-C18 carboxylic acid or its salt, and wherein the incubation is conducted at a temperature ranging from about 30°C to about 35°C for a period of time ranging from about 15 minutes to 24 hours and (3) subjecting the conjugate produced in step (2) to a chromatographic separation process to separate monomeric calicheamicin derivative/antibody conjugates with a loading in the range of 4-10% by weight calicheamicin and with low conjugated fraction (LCF) below 10 percent from unconjugated antibody, calicheamicin derivative, and aggregated conjugates. Further disclosed are compositons with reduced low conjugated fraction (LCF) below 10 percent comprising monomeric calicheamicin derivative/anti-CD22 antibody conjugates having the formula, Pr(-X-W)m wherein: Pr is an anti-CD22 antibody X is a hydrolyzable linker having the formula (CO-Alk1-sp1-Ar-sp2-Alk2-C(Z1)=Q-Sp) as defined herein.
