US 11,999,980 B2
Non-specific nucleases from the genus Pseudomonas for use in facilitating flow of cells through a microfluidic channel
Skander Elleuche, Hamburg (DE); Sarah Schmitz, Cologne (DE); Stefan Miltenyi, Bergisch Gladbach (DE); and Volker Nolle, Kurten (DE)
Assigned to Miltenyi Biotec B.V. & Co. KG, Bergisch Gladbach (DE)
Appl. No. 17/264,748
Filed by Miltenyi Biotec B.V. & Co. KG, Bergisch Gladbach (DE)
PCT Filed Jul. 26, 2019, PCT No. PCT/EP2019/070148
§ 371(c)(1), (2) Date Jan. 29, 2021,
PCT Pub. No. WO2020/025468, PCT Pub. Date Feb. 6, 2020.
Claims priority of application No. 18186232 (EP), filed on Jul. 30, 2018.
Prior Publication US 2021/0301273 A1, Sep. 30, 2021
Int. Cl. C12N 9/22 (2006.01); C12N 9/96 (2006.01); G01N 1/40 (2006.01)
CPC C12N 9/22 (2013.01) [C12N 9/96 (2013.01); G01N 1/4044 (2013.01)] 15 Claims
 
1. A method of preparing a cell suspension, comprising:
(a) obtaining a recombinant nuclease,
(b) incubating the cell suspension with the recombinant nuclease until nucleic acids present in the suspension and/or released from cells are removed, and then
(c) flowing the cells through a microfluidic channel;
wherein the recombinant nuclease comprises an amino acid sequence that is at least 90% identical to the amino acid sequence set forth in SEQ ID NO:2,
wherein the recombinant nuclease when present in a solution at 4° C. has non-specific nuclease (NSN) activity with a kcat value of at least 10 s−1, and
wherein removing nucleic acids from the suspension in step (b) prevents nucleic acids from clogging the microfluidic channel in step (c).