Methods for antibody drug conjugation, purification, and formulation

Abstract

Methods of producing, purifying, and formulating antibody drug conjugates (ADCs) are provided herein. The methods use continuous conjugation processes, single-pass tangential flow filtration, countercurrent diafiltration, and/or in-line process automation technologies. The methods decrease process times and costs and improve product consistency.

1 - 28 . (canceled) 
     
     
         29 . A method for concentrating an antibody drug conjugate (ADC), purifying an ADC, or transferring an ADC to a formulation buffer comprising using single-pass tangential flow filtration and/or countercurrent diafiltration. 
     
     
         30 - 31 . (canceled) 
     
     
         32 . The method of  claim 29 , wherein the ADC was produced in a batch conjugation process. 
     
     
         33 . The method of  claim 29 , wherein the ADC was produced in a continuous conjugation process. 
     
     
         34 . The method of  claim 29 , and wherein the single-pass tangential flow filtration uses an ultrafiltration membrane. 
     
     
         35 . The method of  claim 29 , wherein the single-pass tangential flow filtration uses a diafiltration membrane. 
     
     
         36 . The method of  claim 29 , wherein the method improves the consistency of the ADC production. 
     
     
         37 . The method of  claim 29 , wherein the method decreases the time for ADC production. 
     
     
         38 . The method of  claim 29 , wherein the single-pass tangential flow filtration and/or the countercurrent diafiltration improves the consistency of the ADC production. 
     
     
         39 . The method of  claim 29 , wherein the single-pass tangential flow filtration and/or the countercurrent diafiltration decreases the time for ADC concentration, purification, or transfer. 
     
     
         40 . The method of  claim 29 , wherein the single-pass tangential flow filtration and/or the countercurrent diafiltration decreases the amount of buffer used. 
     
     
         41 . The method of  claim 29 , wherein the method further comprises in-line monitoring of an analyte. 
     
     
         42 . A method for producing an antibody drug conjugate (ADC) wherein (i) in-line monitoring is used to measure the addition of a component to an ADC conjugation reaction, (ii) in-line monitoring of an analyte is used to determine when to stop adding conjugation buffer to an ADC conjugation reaction (iii) in-line monitoring of an analyte is used to determine when to stop recirculating conjugation buffer in an ADC conjugation reaction, (iv) in-line monitoring of an analyte is used to determine when to start rinsing conjugation buffer from an ADC conjugation reaction, and/or (v) in-line monitoring of an analyte is used to determine when to stop adding a conjugation reaction component to an ADC conjugation reaction. 
     
     
         43 - 49 . (canceled) 
     
     
         50 . A method for purifying an antibody drug conjugate (ADC) after an ADC conjugation process wherein the method comprises in-line monitoring of an analyte. 
     
     
         51 .- 77 . (canceled) 
     
     
         78 . The method of  claim 29 , wherein the ADC comprises an antibody or antigen-binding fragment that specifically binds to CD37, CD33, FOLR1, CD123, CD19, cMET, ADAM9, or HER2. 
     
     
         79 .- 83 . (canceled) 
     
     
         84 . The method of  claim 29 , wherein the ADC comprises a linker selected from the group consisting of SMCC, sSPDB, and a peptide linker. 
     
     
         85 . The method of  claim 29 , wherein the ADC comprises a maytansinoid or an indolino-benzodiazepine. 
     
     
         86 - 87 . (canceled) 
     
     
         88 . The method of  claim 29 , wherein the ADC is IMGN529, IMGN779, IMGN853, IMGN632, or Kadcyla. 
     
     
         89 .- 92 . (canceled) 
     
     
         93 . A method of producing an ADC comprising (i) increasing a first temperature of a conjugation process by at least 5° C. to an elevated temperature, (ii) decreasing a first temperature of conjugation process by at least 5° C. to a reduced temperature, (iii) changing a first pH of a conjugation process by at least 0.5 to an altered pH, wherein the elevated temperature, the reduced temperature, or the altered pH is maintained for no more than 20 minutes. 
     
     
         94 .- 139 . (canceled) 
     
     
         140 . An ADC produced according to the method of  claim 29 .