MEMBRANE RUPTURE COMPOSITIONS AND METHODS OF MAKING AND USING SAME

1 . A membrane rupture solution comprising:

one or more purified non-ionic detergents, wherein the non-ionic detergent is selected from the group consisting of ethoxylated alcohol, sorbiton ethoxylate, triblock copolymer and combinations thereof, wherein the detergent is in a concentration range from about 0.01% (w/v) to about 2.0% (w/v); and

a scavenger, in a concentration in a range of 0% to about 2.0%.

2 . The solution of claim 1 , wherein the ethoxylated alcohol is TDA9, tergitol 15-S-9, or combinations thereof.

3 . The solution of claim 1 , wherein the sorbiton ethoxylate is polysorbate 20, polysorbate 80, or combinations thereof.

4 . The solution of claim 1 , wherein the triblock copolymer is poloxamer 188, poloxamer 407, poloxamer 305, or combinations thereof.

5 . The solution of claim 1 , wherein the non-ionic detergent is in a concentration of about 0.25% (w/v).

6 . The solution of claim 1 , wherein the non-ionic detergent is TDA9 in a concentration of about 0.05% (w/v) and polysorbate 20 in a concentration of about 0.2% (w/v).

7 . The solution of claim 1 , wherein the scavenger is selected from the group consisting of ethylenediaminetetraacetic acid (EDTA), ethanol, deoxythymidine triphosphate (DTTP), polyethylenimine (PEI), a PEI derivative, and combinations thereof.

8 . The solution of claim 1 , wherein the non-ionic detergent comprises ethoxylated alcohol at a concentration of about 0.05% (w/v), polysorbate 20 at a concentration of about 0.2% (w/v) and poloxamer 188 at a concentration of about 0.005% (w/v).

9 . The solution of claim 8 , further comprising the scavenger is PEI at a concentration of about 0.1% (w/v).

10 . The solution of claim 1 , further comprising a stabilizer.

11 . The solution of claim 10 wherein the stabilizer is selected from the group consisting of sucrose, magnesium chloride, sodium chloride, PEG of various chain lengths, and combinations thereof.

12 . A method of extracting intracellular material from cells comprising contacting the cells with the membrane rupture solution according to claim 1 , wherein the cells are lysed.

13 . The method according to claim 12 wherein the intracellular material is viral vector, protein and/or peptide.

14 . The method according to claim 12 wherein the cells are mammalian or insect cells.

15 . The method according to claim 12 wherein the solution is directly added to the cells in cell culture media or to a cell suspension, and wherein the cells are then centrifuged to remove cell media and resuspended in a suitable buffer.

16 . The method of claim 15 wherein the buffer is selected from the group consisting of Tris, phosphate, histidine, citrate, acetate, and combinations thereof.

17 . The method of claim 12 , wherein the scavenger is added only after cell lysis.

18 . A method of inactivating viruses comprising contacting a biological product with the membrane rupture solution according to claim 1 .

19 . The method according to claim 18 wherein the biological product contains at least one enveloped virus.

20 . The method according to claim 18 wherein the biological product is exposed to the membrane rupture solution for about 1 minute to about 120 minutes at 2° C. to 40° C.

21 . A method of purifying TDA9 to reduce product-related impurities, comprising:

(1) heating raw material to 50-180° C. while mixing with an inert gas or applying vacuum, wherein the raw material comprises impurities and TDA9;

(2) cooling the material of step (1) to 15-40° C. while mixing with inert gas;

(3) adding 10-50 volume percent of water to the material of step (2) to form a homologized solution;

(4) heating the solution of step (3) to 40-120° C. to complete hydrolysis of volatile impurities; and

(5) filtering the solution of step (4) with a 0.2-1.0 μm filter,

wherein the TDA9 is purified.

22 . The method of claim 21 wherein the impurity comprises ethylene oxide and/or polyethylene glycol.