	US 12,404,502 B2
	Hybrid yeast cell lines for high level production of recombinant protein
Lawrence Forman, San Mateo, CA (US)
Assigned to CHO Plus, Inc.
Filed by CHO Plus, Inc., San Mateo, CA (US)
Filed on Apr. 15, 2022, as Appl. No. 17/721,930.
Application 17/721,930 is a continuation of application No. 16/440,776, filed on Jun. 13, 2019, granted, now 11,649,449.
Application 16/440,776 is a continuation of application No. PCT/US2019/036379, filed on Jun. 10, 2019.
Application 16/440,776 is a continuation of application No. 15/254,852, filed on Sep. 1, 2016, granted, now 10,329,594, issued on Jun. 25, 2019.
Claims priority of provisional application 62/213,880, filed on Sep. 3, 2015.
Prior Publication US 2022/0243192 A1, Aug. 4, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 15/02 (2006.01); C12N 5/071 (2010.01); C12N 5/16 (2006.01); C12N 15/10 (2006.01)

CPC C12N 15/02 (2013.01) [C12N 5/0682 (2013.01); C12N 5/16 (2013.01); C12N 15/10 (2013.01)]

10 Claims

1. A method of obtaining a hybrid yeast cell line adapted for high-level production of protein, comprising:

(a) fusing together and culturing individual cells in a starting population of yeast cells whereby a plurality of cell hybrids is formed, each comprising two or more of said individual cells; then

(b) sorting the cell hybrids according to density of endoplasmic reticulum and/or Golgi apparatus per cell hybrid; and

(c) selecting and recovering cell hybrids that have a relatively high density of endoplasmic reticulum and/or Golgi apparatus per cell hybrid compared with other cell hybrids sorted in step (b);

thereby obtaining a yeast cell line that supports increased production and/or secretion of protein compared with parental cells in the starting population.