Title:
PREPARATION METHOD FOR AND USE OF NK CELL OVER-EXPRESSING CD16A
Document Type and Number:
WIPO Patent Application WO/2023/217182
Kind Code:
A1
Abstract:
Provided in the present application is a method for preparing an NK cell over-expressing CD16a, comprising the following steps: (1) constructing a lentiviral transfection system having a target plasmid of CD16a; (2) culturing HEK 293T cells; (3) uniformly mixing a transfection reagent with the plasmid; (5) packaging lentiviruses; (6) collecting, filtering and concentrating the virus solution; (7) transfecting NK cells resuspended by a second cell culture medium, adding polybrene, and preheating the mixture; (8) centrifuging the mixture; (9) adding a second cell culture medium containing polybrene, uniformly mixing same, and continuing to culture same using a third cell culture medium; and (11) taking out cells, performing flow cytometry to detect the positive rate, performing fluorescence-activated cell sorting to screen out positive cells, inoculating a corresponding system, continuing to proliferate and culture same, and constructing a library. The NK cells over-expressing CD16a prepared by the method of the present application have a high positive rate and a high cell survival rate, can still stably over-express CD16a after multiple passages, and have a good killing performance on cancer cells.
Inventors:
ZENG HAOYU (CN)
SHEN ZHENBO (CN)
JIANG BIYU (CN)
WEI JIAQI (CN)
LIU KANGZHI (CN)
SHEN ZHENBO (CN)
JIANG BIYU (CN)
WEI JIAQI (CN)
LIU KANGZHI (CN)
Application Number:
PCT/CN2023/093245
Publication Date:
November 16, 2023
Filing Date:
May 10, 2023
Export Citation:
Assignee:
GUANGDONG PROCAPZOOM BIOSCIENCES CO LTD (CN)
International Classes:
C12N15/867; C12N5/10
Foreign References:
| CN114807237A | 2022-07-29 | |||
| CN109844099A | 2019-06-04 | |||
| CN112813031A | 2021-05-18 | |||
| US20220073878A1 | 2022-03-10 |
Other References:
HULLSIEK ROBERT, LI YUNFANG, SNYDER KRISTIN M., WANG SAM, DI DA, BORGATTI ANTONELLA, LEE CHAE, MOORE PETER F., ZHU CONG, FATTORI C: "Examination of IgG Fc Receptor CD16A and CD64 Expression by Canine Leukocytes and Their ADCC Activity in Engineered NK Cells", FRONTIERS IN IMMUNOLOGY, FRONTIERS MEDIA, LAUSANNE, CH, vol. 13, 24 February 2022 (2022-02-24), Lausanne, CH , pages 841859, XP093107324, ISSN: 1664-3224, DOI: 10.3389/fimmu.2022.841859
XUE-LIAN LI, YANG BEN-YAN-ZI; FAN ZHAO-XIN: "The Effect of CD16 Monoclonal Antibody and Cytokines on NK Cell Proliferation and Killing Efficiency against K562", BIOLOGICAL CHEMICAL ENGINEERING, vol. 3, no. 4, 28 August 2017 (2017-08-28), pages 30 - 33, 36, XP093107329
XUE-LIAN LI, YANG BEN-YAN-ZI; FAN ZHAO-XIN: "The Effect of CD16 Monoclonal Antibody and Cytokines on NK Cell Proliferation and Killing Efficiency against K562", BIOLOGICAL CHEMICAL ENGINEERING, vol. 3, no. 4, 28 August 2017 (2017-08-28), pages 30 - 33, 36, XP093107329
Attorney, Agent or Firm:
SCIHEAD IP LAW FIRM et al. (CN)
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