Process for manufacturing conjugates of improved homogeneity

• Main Author: JIN SHENGJIN
• Format: Patent
• Language: English
• Published: 29.01.2016
• Subjects: HUMAN NECESSITIES; HYGIENE; MEDICAL OR VETERINARY SCIENCE; PREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES; SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS ORMEDICINAL PREPARATIONS

Discloses a process for preparing a conjugate comprising a cell-binding agent chemically coupled to a maytansinoid, which process comprises: (a) contacting a cell-binding agent with a bifunctional crosslinking reagent in a solution having a pH of about 7.5 to about 9 at a temperature of about -10°C to about 15°C to covalently attach a linker to the cell-binding agent and thereby prepare a first mixture comprising cell-binding agents having linkers bound thereto; (b) conjugating a maytansinoid to the cell-binding agents having linkers bound thereto in the first mixture by reacting the cell-binding agents having linkers bound thereto with a maytansinoid in a solution having a pH of about 4 to about 9 to prepare a second mixture comprising (i) cell-binding agent chemically coupled through the linker to the maytansinoid, (ii) free maytansinoid, and (iii) reaction by-products; and (c) subjecting the second mixture to tangential flow filtration, selective precipitation, non-adsorptive chromatography, adsorptive filtration, adsorptive chromatography, or a combination thereof, to purify the cell binding agents chemically coupled through the linkers to the maytansinoid from the other components of the second mixture and thereby prepare a purified second mixture of cell binding agents chemically coupled through the linkers to the maytansinoid.