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ADCC Enhanced Cell Line Development

Proprietary CHOK1-ADCC+ cell line system

Enhance ADCC activity

CHOK1-ADCC+ Host Cell System

Antibody-dependent cellular cytotoxicity (ADCC), is a mechanism of cell-mediated immune defense whereby an effector cell of the immune system actively lyses a target cell. It is important in the efficacy of cancer antibodies, but with many approved cancer antibodies there is less ADCC than could be desired due to nonspecific IgG competing with the drugs for binding to FcγIIIa (CD16a) on natural killer cells (NK cells). Afucosylated monoclonal antibodies overcome this problem through improved FcγIIIa binding.

The scientists at ProBio developed ADCC enhanced host cell line (CHOK1-ADCC+), which can produce afucosylated monoclonal antibodies with enhanced ADCC activity.

Background of CHOK1-ADCC+ cell line system

The typical ADCC involves activation of NK cells by antibodies in a multi-tiered progression of immune control. Once the Fc receptor of NK cell binds to the Fc region of the antibody, the NK cell releases cytotoxic factors that cause the death of the target cell.

The binding ability of the Fc and FcγRIIIa affects the ADCC activity. Therefore, by enhancing the affinity of the antibody to CD16a, the ADCC activity is enhanced, and the clinical therapeutic effect of the antibody is enhanced.

ProBio scientists strategically knocked out a gene in CHOK1 cells that is responsible for the fucose bind to N-glycans in order to produce afucosylated antibodies.

The antibody produced by modified cell line CHOK1-ADCC+ is successfully afucosylated. CHOK1-ADCC+has a similar performance in antibody expression and growth compared to wild type CHOK1-GenS.

Glycan Profiling
Cell Growth Performance

Features of CHOK1-ADCC+ cell line system

ADCC activity enhanced
- Enhance affinity to FcγIIIa
- Enhance ADCC activity
Service
- Proprietary CHOK1-ADCC+ cell line system to produce afucosylated antibodies
- Transient Expression/Cell pool/Cell line for different customers in late discovery stage or in CMC stage
Host cell System
- 2 CHO-K1 host cell license to choose :
  wild-type CHOK1-GenS
  ADCC enhanced CHOK1-ADCC+
- Purchase in combination with wild type CHOK1-GenS cell line, and get a discounted price.
- Royalty-free
- Direct sublicense from ProBio

Case Study of CHOK1-ADCC+ cell line system

CD16a 158V/F affinity (BLI)
- The affinity to CD16a (158V and 158F) was increased by 3-4 times.
PBMC-based ADCC
- Cytotoxicity is increased by about 10%.
- Effect concentration (EC50) was reduced by 6 times.
- ADCC effect was significantly enhanced.

Service Package of CHOK1-ADCC+ cell line system

Transient Expression using CHOK1-ADCC+

PreCLD Cell Pool Development using CHOK1-ADCC+

ProCLD Cell Line Development using CHOK1-ADCC+

Service Package	Antibody Transient Expression	Cell pool development & developability assessment	Cell line development PCB & PCB stability
Timeline	8.5 weeks	12 weeks (mAb, protein) 14 weeks (bsAb)	16 weeks (mAb, protein) 18 weeks (bsAb)
Delivery	ROA	4 Mix pools	6 PCBs
Application scenario	Late discovery stage, after get human or humanized antibody candidates	Late discovery stage, after get human or humanized antibody candidates	Preclinical CMC development
Benefits to clients	Shorten the timeline, and quickly preview the ADCC activity of antibodies Comprehensive bio analytical methods for ADCC performance evaluation	Evaluate the developability of drug candidate in the same host cell and vector system as CMC Help client to identify the possible risk occurred in the process development The cell clone can be further developed to stable cell line	Proprietary CHOK1-ADCC+ cell line system to produce afucosylated antibodies Guaranteed 60 generation stability Antibody expression levels increase significantly compared to transient expression and cell pool development Compliant for IND filing requirements A variety of detection options are available

Reference:

1.Hashimoto, G.; Wright, P. F.; Karzon, D. T. (1983-11-01). "Antibody-dependent cell-mediated cytotoxicity against influenza virus-infected cells". The Journal of Infectious Diseases. 148 (5): 785–794.

2.Lo Nigro, Cristiana; Macagno, Marco; Sangiolo, Dario; Bertolaccini, Luca; Aglietta, Massimo; Merlano, Marco Carlo (March 2019). "NK-mediated antibody-dependent cell-mediated cytotoxicity in solid tumors: biological evidence and clinical perspectives". Annals of Translational Medicine. 7 (5): 105.