Method and system for providing real-time, in situ biomanufacturing process monitoring and control in response to IR spectroscopy

Abstract

A method and system for providing real-time, biomanufacturing process monitoring and control in response to infra-red (IR) spectroscopic fingerprinting of a biomolecule. IR spectroscopy is used to fingerprint an active biomolecule in situ in a biomanufacturing process. In one embodiment, Fourier Transform Infra-red spectroscopy (FTIR) is used to determine whether an active or aged biomolecule is present in stages of a biomanufacturing process. In one preferred example, the biomanufacturing process manufactures a biomaterial in bulk. The biomanufacturing process has four stages: bioproduction, recovery, purification, and bulk storage. FTIR spectroscopy is used to monitor the optimization of each process step by providing feedback controls, and to fingerprint in real-time, in situ whether active biomolecules are present in each stage.

What is claimed is:  
     
         1 . A method for controlling a biomanufacturing process for a biomolecule, comprising the steps of: 
 enabling detection of an infra-red spectra in situ and in real-time during at least one stage of said biomanufacturing process; and    generating at least one control signal in response to the detected infra-red spectra, wherein said control signal enables a control step in said biomanufacturing process.    
     
     
         2 . The method of  claim 1 , wherein said biomanufacturing process comprises a bioproduction stage.  
     
     
         3 . The method of  claim 2 , further comprising a recovery stage.  
     
     
         4 . The method of  claim 3 , further comprising a purification stage.  
     
     
         5 . The method of  claim 4 , further comprising a bulk formulation and storage stage.  
     
     
         6 . The method of  claim 2 , wherein said bioproduction stage comprises a process selected from the group consisting of fermentation, cell culture, semisynthetic bioprocessing, extractive processes, catabolic processes, and production of therapeutic viruses.  
     
     
         7 . The method of  claim 2 , wherein said bioproduction stage comprises a fermentation reaction, and wherein homeostasis of said fermentation reaction is maintained by: 
 enabling detection of an infra-red spectra of said fermentation in sits and in real-time, and    generating at least one control signal in response said infra-red spectra; whereby said fermentation conditions are optimized.    
     
     
         8 . The method of  claim 7 , wherein said control signal enables an adjustment of the conditions of said fermentation reaction, said adjustment being selected from the group consisting of a glucose feed adjustment, a gas feed adjustment, a salts and vitamin feed adjustment, an ammonium hydroxide adjustment, a temperature adjustment, and an agitation speed adjustment.  
     
     
         9 . The method of  claim 3 , wherein said recovery stage comprises at least one recovery step.  
     
     
         10 . The method of  claim 9 , wherein said recovery step is selected from the group consisting of: cell breakage, extraction, denaturation, refolding, oxidation, diafiltration, lyophilization, and concentration.  
     
     
         11 . The method of  claim 9 , wherein said recovery step comprises a refolding reaction.  
     
     
         12 . The method of  claim 11 , comprising the steps of: 
 enabling detection of an infra-red spectra of said biomolecule in situ and in real-time during said refolding reaction; and    generating at least one control signal in response to said infra-red spectra, wherein said control signal enables a recovery control step in said refolding reaction.    
     
     
         13 . The method of  claim 12 , wherein said recovery control step comprises regulation of the rate of oxidation in said refolding reaction.  
     
     
         14 . The method of  claim 13 , wherein said control signal enables regulation of a gas feed to said refolding reaction.  
     
     
         15 . The method of  claim 4 , wherein said purification stage comprises at least one purification step.  
     
     
         16 . The method of  claim 15 , wherein said purification step comprises a process selected from the group consisting of precipitation, continuous sucrose gradient centrifugation, filtration, electrophoresis, and chromatographic separation.  
     
     
         17 . The method of  claim 16 , wherein said chromatographic separation process comprises a step selected from the group consisting of anion exchange chromatography, cation exchange chromatography, gel filtration chromatography, hydrophobic interaction chromatography, chromatofocusing, hydroxylapatite chromatography, and affinity chromatography.  
     
     
         18 . The method of  claim 17 , comprising the steps of: 
 enabling detection of an infra-red spectra of the flow-through from said chromatography step in situ and in real-time; and    generating at least one control signal in response to said infra-red spectra, wherein said control signal enables at least one flow-through control step in said chromatography step.    
     
     
         19 . The method of  claim 18 , wherein said flow-through control step comprises enabling a shunt to allow repetition of said chromatography step.  
     
     
         20 . The method of  claim 18 , comprising the steps of: 
 enabling detection of an infra-red spectra of the eluate from said chromatography step in situ and in real time; and    generating at least one control signal in response to said infra-red spectra, wherein said control signal enables at least one elution control step in said chromatography step.    
     
     
         21 . The method of  claim 20 , wherein said elution control step comprises enabling an adjustment to said chromatography step selected from the group consisting of adjustment of the elution gradient curve, adjustment of the flow rate, adjustment of the temperature, and adjustment of the flow rate.  
     
     
         22 . A method for controlling a biomanufacturing process for a biomolecule, comprising the steps of: 
 detecting an infra-red spectra in situ and in real-time during at least one stage of said biomanufacturing process; and    generating at least one control signal in response to the detected infra-red spectra, wherein said control signal enables a control step in said biomanufacturing process.    
     
     
         23 . The method of  claim 22 , wherein said biomanufacturing process comprises a bioproduction stage.  
     
     
         24 . The method of  claim 23 , further comprising a recovery stage.  
     
     
         25 . The method of  claim 23 , further comprising a purification stage.  
     
     
         26 . The method of  claim 23 , further comprising a bulk formulation and storage stage.  
     
     
         27 . A method for monitoring in real-time a biomolecule formulated in a storage solution placed in a bulk container, comprising the steps of: 
 detecting an infra-red spectra of the biomolecule in situ and in real time; and    generating a bioactivity indication signal in response to the detected infra-red spectra, wherein the bioactivity indication signal provides an indication of the degradation of the biomolecule during storage.    
     
     
         28 . The method of  claim 27 , further comprising the steps of: 
 generating at least one control signal in response to the detected infra-red spectra, wherein the control signal regulates a condition of an environment of the bulk container to maintain a predetermined optimum condition.    
     
     
         29 . The method of  claim 28 , wherein the control signal regulates at least one of a temperature of the bulk container and a nitrogen concentration in a headspace in the bulk container.  
     
     
         30 . The method of  claim 27 , further comprising the steps of: 
 measuring at least one environment condition of the bulk container; and    generating at least one control signal in response to each measured condition, wherein the control signal regulates the respective measured condition of an environment of the bulk container to maintain a predetermined optimum condition.    
     
     
         31 . The method of  claim 27 , further comprising the steps of: 
 measuring at least one environment condition of the bulk container; and    generating at least one environment condition indication signal in response to each measured condition, wherein each environment condition indication signal provides a current status condition of a condition of an environment of the bulk container.    
     
     
         32 . The method of  claim 31 , wherein said measuring at least one environment condition of the bulk container step comprises measuring at least one of particle size, UV radiation, oxygen concentration, pH level, conductivity, and temperature.  
     
     
         33 . A system for monitoring in real-time a biomolecule formulated in a storage solution placed in a bulk container, comprising: 
 an FTIR spectrometer, coupled to that bulk container, that detects an infra-red spectra of the biomolecule in situ and in real time, and generates a bioactivity indication signal in response to the detected infra-red spectra, wherein the bioactivity indication signal provides an indication of the degradation of the biomolecule during storage.    
     
     
         34 . The system of  claim 33 , further comprising: 
 means for measuring at least one environment condition of the bulk container; and    means for generating at least one environment condition indication signal in response to each measured condition, wherein each environment condition indication signal provides a current status of a condition of an environment of the bulk container.    
     
     
         35 . A method for automated, accelerated storage studies of a biomolecule in an array of wells in real-time, comprising the steps of: 
 performing FTIR measurements of the biomolecule in the array of wells at varying temperatures;    determining bioactivity ratios based on said FTIR measurements;    generating a multi-dimensional map of stability parameters including the bioactivity ratios; and    finding areas of stability parameters associated with maximum stability.    
     
     
         36 . The method of  claim 35 , further comprising the step of outputting the areas of stability parameters associated with maximum stability.  
     
     
         37 . A system for automated, accelerated storage studies of a biomolecule in an array of wells in real-time, comprising: 
 an accelerated storage analyzer; and    a FTIR spectrometer that performs FTIR measurements of the biomolecule in the array of wells at varying temperatures in response to signals from said accelerated storage analyzer; 
 wherein said accelerated storage analyzer determines bioactivity ratios based on said FTIR measurements, generates a multi-dimensional map of stability parameters including the bioactivity ratios, and finds areas of stability parameters associated with maximum or near maximum stability.  
   
     
     
         38 . The system of  claim 37 , wherein said accelerated storage analyzer outputs the areas of stability parameters associated with maximum or near maximum stability to a display or storage device.