6YSC

GLYCOSYLATED KNOB-HOLE/DUMMY FC FRAGMENT

Summary for 6YSC

• Entry DOI: 10.2210/pdb6ysc/pdb
• Descriptor: Immunoglobulin gamma-1 heavy chain, beta-D-galactopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-2)-alpha-D-mannopyranose-(1-6)-[alpha-D-mannopyranose-(1-3)]beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-[alpha-L-fucopyranose-(1-6)]2-acetamido-2-deoxy-beta-D-glucopyranose, ... (4 entities in total)
• Functional Keywords: bispecific antibody, fc engineering, knob-into-hole, force, format chain exchange, immune system
• Biological source: Homo sapiens (Human); More
• Total number of polymer chains: 2
• Total formula weight: 55511.19

Authors

Kuglstatter, A.,Leibrock, L.,Benz, J. (deposition date: 2020-04-22, release date: 2020-10-14, Last modification date: 2024-11-06)

Primary citation

Dengl, S.,Mayer, K.,Bormann, F.,Duerr, H.,Hoffmann, E.,Nussbaum, B.,Tischler, M.,Wagner, M.,Kuglstatter, A.,Leibrock, L.,Buldun, C.,Georges, G.,Brinkmann, U.
Format chain exchange (FORCE) for high-throughput generation of bispecific antibodies in combinatorial binder-format matrices.
Nat Commun, 11:4974-4974, 2020

PubMed Abstract: Generation of bispecific antibodies (bsAbs) requires a combination of compatible binders in formats that support desired functionalities. Here, we report that bsAb-matrices can be generated by Format Chain Exchange (FORCE), enabling screening of combinatorial binder/format spaces. Input molecules for generation of bi/multi-valent bsAbs are monospecific entities similar to knob-into-hole half-antibodies, yet with complementary CH3-interface-modulated and affinity-tagged dummy-chains. These contain mutations that lead to limited interface repulsions without compromising expression or biophysical properties of educts. Mild reduction of combinations of educts triggers spontaneous chain-exchange reactions driven by partially flawed CH3-educt interfaces resolving to perfect complementarity. This generates large bsAb matrices harboring different binders in multiple formats. Benign biophysical properties and good expression yields of educts, combined with simplicity of purification enables process automation. Examples that demonstrate the relevance of screening binder/format combinations are provided as a matrix of bsAbs that simultaneously bind Her1/Her2 and DR5 without encountering binder or format-inflicted interferences.

PubMed: 33009381
DOI: 10.1038/s41467-020-18477-7

Experimental method

X-RAY DIFFRACTION (2.05 Å)