NKp46-specific single domain antibodies enable facile engineering of various potent NK cell engager formats

被引：15

作者：

Lipinski, Britta ^{[1
,2
]}

Arras, Paul ^{[1
]}

Pekar, Lukas ^{[1
]}

Klewinghaus, Daniel ^{[1
]}

Boje, Ammelie Svea ^{[3
,4
]}

Krah, Simon ^{[1
]}

Zimmermann, Jasmin ^{[1
,2
]}

Klausz, Katja ^{[3
,4
]}

Peipp, Matthias ^{[3
,4
]}

Siegmund, Vanessa ^{[5
]}

Evers, Andreas ^{[6
]}

Zielonka, Stefan ^{[1
,2
,7
]}

机构：

[1] Merck Healthcare KGaA, Prot Engn & Antibody Technol, Darmstadt, Germany

[2] Tech Univ Darmstadt, Inst Organ Chem & Biochem, Darmstadt, Germany

[3] Univ Hosp Schleswig Holstein, Dept Internal Med 2, Div Antibody Based Immunotherapy, Kiel, Germany

[4] Christian Albrechts Univ Kiel, Kiel, Germany

[5] Merck Healthcare KGaA, Prot & Cell Sci, Darmstadt, Germany

[6] Merck Healthcare KGaA, Computat Chem & Biol, Darmstadt, Germany

[7] Merck Healthcare KGaA, Prot Engn & Antibody Technol, Frankfurter Str 250, D-64293 Darmstadt, Germany

来源：

PROTEIN SCIENCE | 2023年 / 32卷 / 03期

关键词：

ADCC; antibody engineering; bispecific antibody; multifunctional antibody; NK cell engager; NK cell redirection; NKp46; single domain antibody; valencies; VHH; FC-GAMMA-RIIA; MELANOMA-CELLS; RECEPTOR; NKP30; LIGAND; EGFR; POLYMORPHISMS; CYTOTOXICITY; IMPACT;

D O I：

10.1002/pro.4593

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Herein, we describe the generation of potent NK cell engagers (NKCEs) based on single domain antibodies (sdAbs) specific for NKp46 harboring the humanized Fab version of Cetuximab for tumor targeting. After immunization of camelids, a plethora of different VHH domains were retrieved by yeast surface display. Upon reformatting into Fc effector-silenced NKCEs targeting NKp46 and EGFR in a strictly monovalent fashion, the resulting bispecific antibodies elicited potent NK cell-mediated killing of EGFR-overexpressing tumor cells with potencies (EC(50)killing) in the picomolar range. This was further augmented via co-engagement of Fc gamma receptor IIIa (Fc gamma RIIIa). Importantly, NKp46-specific sdAbs enabled the construction of various NKCE formats with different geometries and valencies which displayed favorable biophysical and biochemical properties without further optimization. By this means, killing capacities were further improved significantly. Hence, NKp46-specific sdAbs are versatile building blocks for the construction of different NKCE formats.

引用

页数：16

共 62 条

[31] Grabbing the Bull by Both Horns: Bovine Ultralong CDR-H3 Paratopes Enable Engineering of 'Almost Natural' Common Light Chain Bispecific Antibodies Suitable For Effector Cell Redirection [J].