Patent ID: 11958881
Assignee: ZHEJIANG UNIVERSITY
Field: Chemical engineering (Chemistry)
Classification: CPC C  B | IPC B  C

Claim 0:
1. An optimization method for capturing proteins by a multi-column continuous chromatography (MCC), wherein the MCC is used for a protein capture, and a number of columns is greater than or equal to 3, the optimization method comprises the following steps:
step 0, loading a target protein containing solution into a chromatographic column of the MCC for continuous capture, wherein the target protein is a monoclonal antibody and the chromatographic column of the MCC comprises a protein A affinity resin;
step 1, under conditions of a set loading protein concentration and an arbitrary load residence time, performing a single time of a protein breakthrough experiment to obtain a protein breakthrough curve;
step 2, under a set breakthrough percentage for a target protein, integrating the protein breakthrough curve to obtain a single-column loading capacity of the MCC as a first formula:, A
  =
  
   
    
     
      ∫
      
       t
       =
       0
      
      
       t
       
        l
        ⁢
        _
        ⁢
        s
       
      
     
     
      
       [
       
        
         c
         exp
        
        -
        
         c
         ⁡
         (
         t
         )
        
       
       ]
      
      ⁢
      dt
     
    
    
     RT
     C
    
   
   -
   
    
     c
     exp
    
    ×
    s
   
  
 

in the first formula, s is the set breakthrough percentage for the target protein, A (g/L) is the single-column loading capacity of the MCC obtained by integrating the protein breakthrough curve at the set breakthrough percentage s, t is a loading time, t1_s (min) is a loading time until reaching the set breakthrough percentage s, and cexp (g/L) is a loading protein concentration; c(t) (g/L) is a breakthrough protein concentration, and RTc (min) is a single-column residence time of an interconnected load of the MCC;
establishing a linear relationship between an interconnected load time tC and a load residence time RTC through the single-column loading capacity further comprises the following step:
substituting the single-column loading capacity obtained in step 2 into a second formula, t
    C
   
   =
   
    
     
      A
      ×
      
       RT
       C
      
     
     
      c
      exp
     
    
    -
    
     t
     CW
    
   
  
  ,, wherein tC (min) is the interconnected load time of the MCC, RTC (min) is the single-column residence time of the interconnected load of the MCC, A (g/L) is the single-column loading capacity of the MCC obtained in step 2 by integrating the protein breakthrough curve at the set breakthrough percentage for the targe protein, tCW (min) is an interconnected wash time of the MCC, and cexp (g/L) is the loading protein concentration; through the first formula and the second formula, the linear relationship between the interconnected load time tc and the load residence time RTC is obtained;
step 3, solving an optimal number of operating columns for capturing the proteins by the MCC under the set loading protein concentration and a set protein breakthrough percentage based on the linear relationship between the interconnected load time tC and the load residence time RTC obtained in step 2 further comprises the following steps:
drawing a line tC-RTC in a t-RT coordinate system based on the linear relationship between the interconnected load time tC and the load residence time RTC obtained in step 2, and drawing a line of a third formula, t
  =
  
   
    
     t
     RR
    
    -
    
     
      (
      
       N
       -
       3
      
      )
     
     ⁢
     
      t
      CW
     
    
   
   
    (
    
     N
     -
     2
    
    ), in the t-RT coordinate system, wherein tCW (min) is the interconnected wash time of the MCC, tRR (min) is a recovery and regeneration (R-R) time of the MCC and comprises a sum of a washing time, an elution time , and a regeneration time, and N is a number of the operating columns; by adjusting a N value, an intersection of two lines is changed so that a load residence time corresponding to the intersection is within a set residence time range; if only one N value meets above conditions, then the N value is the optimal number of the operating columns for capturing the proteins by the MCC under the set loading protein concentration and the set protein breakthrough percentage; if two or more N values meet the above conditions, a largest N value is selected as the optimal number of the operating columns;
step 4, solving an optimal load residence time for the capturing proteins by the MCC under the set loading protein concentration, the set protein breakthrough percentage, and the optimal number of the operating columns based on the linear relationship between the interconnected load time tC and the load residence time RTC obtained in step 2;
step 5, solving a maximum productivity of capturing the proteins by the MCC based on the optimal load residence time obtained in step 4; and
step 6, optimizing the number of operating columns and the load residence time of the MCC based on the maximum productivity obtained in step 5, and continuously capturing the target protein by the MCC having the optimized number of operating columns and load residence time for the target protein.