	US 12,378,526 B2
	Composition for culturing natural killer cells and method for preparing natural killer cells by using same
Myoung Ho Jang, Seoul (KR); Chun-Pyo Hong, Gyeonggi-do (KR); Dong Woo Ko, Seoul (KR); and June Sub Lee, Gyeonggi-do (KR)
Assigned to GI CELL, INC., Gyeonggi-Do (KR)
Filed by GI CELL, INC., Gyeonggi-do (KR)
Filed on May 17, 2022, as Appl. No. 17/746,835.
Application 17/746,835 is a continuation of application No. PCT/KR2020/016376, filed on Nov. 19, 2020.
Claims priority of application No. 10-2019-0149779 (KR), filed on Nov. 20, 2019; and application No. 10-2020-0015802 (KR), filed on Feb. 10, 2020.
Prior Publication US 2022/0372442 A1, Nov. 24, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 5/0783 (2010.01); A61K 35/17 (2015.01); C07K 14/55 (2006.01); C07K 14/705 (2006.01)

CPC C12N 5/0646 (2013.01) [A61K 35/17 (2013.01); C12N 5/0636 (2013.01); C12N 2501/2302 (2013.01); C12N 2501/51 (2013.01)]

10 Claims

1. A method for culturing an isolated natural killer cell, which comprises:

culturing the isolated natural killer cell without CD3+ or CD56− cells in a composition comprising a fusion protein dimer,

wherein the fusion protein comprises the following structural formula (I) or (II):

N′—X-[linker (1)]n-Fc domain-[linker (2)]m-Y—C′— formula (I)

N′—Y-[linker (1)]n-Fc domain-[linker (2)]m-X—C′— formula (II),

wherein, N′ is the N-terminus of the fusion protein,

C′ is the C-terminus of the fusion protein,

X is a CD80 protein or a fragment thereof,

Y is an IL-2 variant thereof,

the linkers (1) and (2) are peptide linkers, and

n and m are each independently 0 or 1,

wherein the CD80 fragment comprises the extracellular domain of CD80,

wherein the IL-2 variant is obtained by any one selected from the following substitution combinations (a) to (d) in the amino acid sequence of SEQ ID NO: 10:

(a) R38A/F42A

(b) R38A/F42A/Y45A

(d) R38A/F42A/L72G.