Affinity-Based Analytical Purification Of Biotherapeutics For Bioprocess Monitoring

Abstract

The invention as disclosed herein provides a method for purifying a non-antibody protein from solution, comprising a chromatography step wherein the solution is passed over an affinity construct containing an affinity ligand-coupled solid support, wherein the affinity construct is associated with a bioprocess unit operation, and isolating the non-antibody protein from solution.

What is claimed is: 
     
         1 . A method for purifying a non-antibody protein (NAP) from a heterogeneous solution, the method comprising:
 a) contacting the heterogeneous solution comprising the NAP with an affinity construct comprising a solid support coupled to an affinity ligand that binds the NAP; and   b) isolating the affinity-purified NAP from the heterogeneous solution;
 wherein the affinity-purified NAP comprises a critical quality attribute (CQA) that is predictive of the CQA of the NAP produced by a multi-step bioprocess. 
   
     
     
         2 . A method of predicting a critical quality attribute (CQA) of a non-antibody protein (NAP) produced by a bioprocess unit operation within a multi-step bioprocess, the method comprising
 a) purifying the affinity-purified NAP from solution according to the method of  claim 1 ;   a) determining the CQA of the affinity-purified NAP;   b) transforming the CQA of the affinity-purified NAP using a predetermined predictive model,
 wherein the predictive model is specific for one or more of the bioprocess unit operations within the multi-step bioprocess; and 
   c) predicting the CQA of 8; the NAP produced by the bioprocess unit operation within a multi-step bioprocess based on the transformed CQA of the affinity-purified NAP.   
     
     
         3 . The method of  claim 2 , wherein the bioprocess unit operation is a bioreactor process, seed train, capture chromatography, intermediate chromatography, filtration, centrifugation, precipitation, flocculation, UV irradiation, or viral inactivation. 
     
     
         4 . A method of monitoring or controlling a bioprocess unit operation within a multi-step bioprocess for producing a NAP, the method comprising:
 a) purifying the affinity-purified NAP from solution according to the method of  claim 1 ;   b) determining a critical quality attribute (CQA) of the affinity-purified NAP;   c) comparing the CQA of the affinity-purified NAP to the CQA of the NAP produced by the bioprocess unit operation within a multi-step bioprocess;   d) calculating a difference between the CQA of the affinity-purified NAP and the NAP produced by the bioprocess unit operation within a multi-step bioprocess; and   e) monitoring or controlling the bioprocess unit operation based on the calculated difference in CQAs.   
     
     
         5 . A method of monitoring or controlling a bioprocess unit operation within a multi-step bioprocess for producing a NAP, the method comprising:
 a) purifying the affinity-purified NAP from solution according to the method of  claim 1 ;   b) determining a critical quality attribute (CQA) of the affinity-purified NAP;   c) transforming the CQA of the affinity-purified NAP using a predetermined predictive model,
 wherein the predictive model is specific for one or more of the bioprocess unit operations within the multi-step bioprocess; and 
   d) monitoring or controlling the bioprocess unit operation based on the predicted CQAs.   
     
     
         6 . A method of developing a bioprocess for a NAP, the method comprising:
 a) processing the NAP using a bioprocess unit operation within the multi-step bioprocess;   b) purifying the affinity-purified NAP from solution according to the method of  claim 1 ;   c) determining a critical quality attribute (CQA) of the NAP produced using the bioprocess unit operation;   d) determining the CQA of the affinity-purified NAP;   e) comparing the CQA of the affinity-purified NAP to the CQA of the NAP produced using the bioprocess unit operation;   f) calculating a difference between the CQA of the affinity-purified NAP and the CQA of the NAP produced using the bioprocess unit operation, and   g) developing the bioprocess by modifying the bioprocess unit operation based on the calculated difference in CQAs.   
     
     
         7 . A method of developing a bioprocess for a NAP, the method comprising:
 a) processing the NAP using a bioprocess unit operation within the bioprocess;   b) purifying the affinity-purified NAP from solution according to the method of  claim 1 ;   c) determining the CQA of the affinity-purified NAP;   d) transforming the CQA of the affinity-purified NAP using a predetermined predictive model,
 wherein the predictive model is specific for one or more of the bioprocess unit operations within the multi-step bioprocess; and 
   e) developing the multi-step bioprocess by modifying the bioprocess unit operation based on the predicted CQAs.   
     
     
         8 . The method of  claim 2 ,  5 , or  7 , wherein the transformed CQA of the affinity-purified NAP is substantially equivalent to the NAP produced by a bioprocess unit operation within the multi-step bioprocess. 
     
     
         9 . The method of  claim 1 , wherein the CQA of the NAP is determined at the following stages within the multi-step bioprocess:
 a) immediately upstream of a particular bioprocess unit operation;   b) immediately downstream of the particular bioprocess unit operation;   c) both upstream and downstream of a particular bioprocess unit operation, or   d) within a particular bioprocess unit operation at one or more timepoints.   
     
     
         10 . The method of  claim 1 , wherein the CQA of the NAP is determined at one or more timepoints within the multi-step bioprocess. 
     
     
         11 . The method of  claim 1 , wherein the bioprocess is selected from the group consisting of continuous, semi-continuous, and batch. 
     
     
         12 . The method of  claim 11 , wherein the CQA is measured using a high-throughput or rapid analytical technique. 
     
     
         13 . The method of  claim 12 , wherein the analytical technique is high-performance liquid chromatography (HPLC), differential refractometry, fluorescence, ultra-performance liquid chromatography (UPLC), multi-angle laser light scattering analysis (MALLS), mass spectroscopy, tandem mass spectroscopy, isoelectric focusing, or differential scanning calorimetry. 
     
     
         14 . The method of  claim 1 , wherein the NAP is a biotherapeutic drug substance or a commercial biologic. 
     
     
         15 . The method of  claim 1 , wherein the multi-step bioprocess for the NAP is a commercial or manufacturing process. 
     
     
         16 . The method of  claim 1 , wherein the CQA is product purity, potency, charged isoform profile, post-translational modifications, oxidation, reductions, deamidation, adduct formation, clipped forms, enzymatic cleavage, specific activity, peptide map, dimer content, product aggregation, site specific glycosylation, total glycans, or glycosylation profile. 
     
     
         17 . The method of  claim 1 , wherein one or more purification steps are performed by a robot. 
     
     
         18 . The method of  claim 1 , wherein the ligand of the affinity ligand-coupled based solid support is an antibody. 
     
     
         19 . The method of  claim 1 , wherein the affinity construct is integrated with the bioprocess in a manner selected from the group consisting of at-line mode, offline mode, and in-line mode. 
     
     
         20 . The method of  claim 1 , wherein the affinity construct comprises a parameter that is optimized to maximize the quality or purity of the affinity-purified NAP.