Process for separating and determining the viral load in a pancreatin sample

1. A process for manufacturing a pharmaceutical composition comprising pancreatin, comprising the steps of:

(a) producing a liquid pancreatin test sample from a quantity of pancreatin, the liquid pancreatin test sample comprising pancreatin, a cell culture medium suitable for cell line used to culture virus, and one or more antibiotics;

(b) centrifugation of the liquid pancreatin test sample at a relative centrifugal force of less than 10,000×g, in which viruses with sedimentation constants of greater than about 120 S do not form a pellet whereby the low-speed centrifugation of the liquid pancreatin test sample produces a supernatant;

(c) ultracentrifugation of the liquid pancreatin test sample supernatant with a discontinuous gradient medium at a relative centrifugal force greater than 100,000×g transfer a virus from the liquid pancreatin test sample supernatant into a target fraction of the discontinuous gradient medium, whereby

the target fraction is quantitatively separated from the liquid pancreatin test sample supernatant;

(d) quantitatively determining a viral load of the pancreatin test sample by culturing the target fraction with a population of cells, assessing the population of cells for viral infection, and calculating a viral titer in the target traction; and

(e) producing a dosage form suitable for oral administration from the quantity of pancreatin when the liquid pancreatin test sample is determined to be substantially free of virus;

wherein steps (a), (b), and (c) are performed without altering the viral load.

2. The process of claim 1 , wherein the target fraction is filtered through a micro filter before quantitatively determining the viral load.

3. The process of claim 1 , wherein the liquid pancreatin test sample suspension is at a temperature of between about 0° C. and about 15° C. during low-speed centrifugation.

4. The process of claim 1 , wherein the centrifugation of step (b) is at a relative centrifugal force between about 1,500×g and about 5,000×g.

5. The process of claim 1 , wherein the centrifugation of step (b) is carried out for longer than five minutes.

6. The process of claim 1 , wherein ultracentrifugation is carded out for a duration of more than one hour.

7. The process of claim 1 , wherein the ultracentrifugation is carried out for a duration of between about 2 hours and about 8 hours and the relative centrifugal force is between about 200,000×g and about 350,000×g.

8. The process of claim 1 , wherein the liquid pancreatin test sample is at a temperature of between about 0° C. and about 15° C. during ultracentrifugation.

9. A process for manufacturing a pharmaceutical composition containing pancreatin, comprising

(a) providing or obtaining a pancreatin test sample from a quantity of pancreatin, wherein the pancreatin test sample further comprises a cell culture medium suitable for a cell line used to culture virus,

(b) isolating a target fraction of the test sample in which transfer of a virus into the target fraction is substantially complete; wherein the isolating step comprises (i) centrifugation of the pancreatin test sample at a relative centrifugal force of less than 10,000×g to produce a pancreatin test sample supernatant, in which viruses with sedimentation constants of greater than about 120 S do not form a pellet: and (ii) ultracentrifugation of the pancreatin test sample supernatant with a discontinuous gradient medium at a relative centrifugal force greater than 100,000×g, whereby the target fraction is quantitatively separated from the pancreatin test sample supernatant;

(c) quantitatively determining a viral load of the pancreatin test sample by culturing the target fraction with a population of cells, assessing the population of cells for viral infection, and calculating a viral titer in the target fraction, and

(d) processing at least a portion of the quantity of pancreatin to obtain the pharmaceutical composition when the pancreatin test sample is determined to be substantially free of virus;

wherein steps (a) and (b) are performed without altering a viral load.

10. The process of claim 9 , wherein the processing step comprises combining the quantity of pancreatin with one or more pharmaceutically acceptable excipients.

11. The process of claim 9 , wherein the processing step comprises preparing pancreatin micropellets.

12. The process of claim 11 , further comprising coating the pancreatin micropellets with a gastric acid resistant coating.

13. The process of claim 12 , wherein the gastric acid resistant coating is hydroxypropyl-methylcellulose acetate succinate (HPMCAS), hydroxypropylmethytceltulose phthalate (HPMCP), cellulose acetate phthalate (CAP) or polyvinyl acetate phthalate (PVAP).

14. The process of claim 12 , wherein the gastric acid resistant coating is hydroxypropylmethylcaulose phthalate (HPMCP).