Humanization of a mouse anti-human complement C6 monoclonal antibody as a potential therapeutic for certain complement-mediated diseases

Lingjun Zhang; Kathryn Armour; Jin Y. Chen; Agathi Mylona; Maojing Yang; Gregers R. Andersen; Jaroslaw P. Maciejewki; Preeti Bakrania; Feng Lin

doi:10.1016/j.molimm.2024.03.010

Humanization of a mouse anti-human complement C6 monoclonal antibody as a potential therapeutic for certain complement-mediated diseases

Lingjun Zhang, Kathryn Armour, Jin Y. Chen, Agathi Mylona, Maojing Yang, Gregers R. Andersen, Jaroslaw P. Maciejewki, Preeti Bakrania, Feng Lin^*

^*Corresponding author for this work

Department of Molecular Biology and Genetics - Protein Science

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

Abstract

The assembly of tissue-damaging membrane attack complexes (MACs; C5b–9) is a major mechanism by which excessive complement activation causes diseases. We previously developed a mouse anti-human C6 monoclonal antibody (mAb) 1C9 that selectively inhibits the assembly of MACs in human and non-human primates. In this project, we found that 1C9 also cross-reacted with rat and guinea pig C6, and determined its binding domains on C6 using different truncated C6 proteins. We then humanized the anti-C6 mAb by molecular modeling and complementarity-determining region grafting. After screening a library of 276 humanized variants with different combinations of humanized light and heavy chains in biophysical assays, we identified clone 3713 with the best developability profile, and an increased affinity against C6 when compared with the parental 1C9 mAb. This humanized 3713 mAb inhibited human, monkey, and rat complement-mediated hemolysis in vitro, and more importantly, it significantly reduced complement-mediated hemolysis in vivo in rats. These results demonstrated the successful humanization of the anti-C6 mAb and suggested that the humanized 3713 mAb could be further developed as a new therapeutic that selectively targets MAC for certain complement-mediated pathological conditions.

Original language	English
Journal	Molecular immunology
Volume	170
Pages (from-to)	19-25
Number of pages	7
ISSN	0161-5890
DOIs	https://doi.org/10.1016/j.molimm.2024.03.010
Publication status	Published - Jun 2024

Keywords

Antibody
C6
Complement
Humanization
Guinea Pigs
Humans
Rats
Cross Reactions/immunology
Antibodies, Monoclonal, Humanized/immunology
Complement Activation/immunology
Animals
Hemolysis/drug effects
Antibodies, Monoclonal/immunology
Complement Membrane Attack Complex/immunology
Mice
Complement C6/immunology

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@article{5420402f8b1b46d89532dfbde845ac05,

title = "Humanization of a mouse anti-human complement C6 monoclonal antibody as a potential therapeutic for certain complement-mediated diseases",

abstract = "The assembly of tissue-damaging membrane attack complexes (MACs; C5b–9) is a major mechanism by which excessive complement activation causes diseases. We previously developed a mouse anti-human C6 monoclonal antibody (mAb) 1C9 that selectively inhibits the assembly of MACs in human and non-human primates. In this project, we found that 1C9 also cross-reacted with rat and guinea pig C6, and determined its binding domains on C6 using different truncated C6 proteins. We then humanized the anti-C6 mAb by molecular modeling and complementarity-determining region grafting. After screening a library of 276 humanized variants with different combinations of humanized light and heavy chains in biophysical assays, we identified clone 3713 with the best developability profile, and an increased affinity against C6 when compared with the parental 1C9 mAb. This humanized 3713 mAb inhibited human, monkey, and rat complement-mediated hemolysis in vitro, and more importantly, it significantly reduced complement-mediated hemolysis in vivo in rats. These results demonstrated the successful humanization of the anti-C6 mAb and suggested that the humanized 3713 mAb could be further developed as a new therapeutic that selectively targets MAC for certain complement-mediated pathological conditions.",

keywords = "Antibody, C6, Complement, Humanization, Guinea Pigs, Humans, Rats, Cross Reactions/immunology, Antibodies, Monoclonal, Humanized/immunology, Complement Activation/immunology, Animals, Hemolysis/drug effects, Antibodies, Monoclonal/immunology, Complement Membrane Attack Complex/immunology, Mice, Complement C6/immunology",

author = "Lingjun Zhang and Kathryn Armour and Chen, {Jin Y.} and Agathi Mylona and Maojing Yang and Andersen, {Gregers R.} and Maciejewki, {Jaroslaw P.} and Preeti Bakrania and Feng Lin",

note = "Publisher Copyright: {\textcopyright} 2024 The Authors",

year = "2024",

month = jun,

doi = "10.1016/j.molimm.2024.03.010",

language = "English",

volume = "170",

pages = "19--25",

journal = "Molecular immunology",

issn = "0161-5890",

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Humanization of a mouse anti-human complement C6 monoclonal antibody as a potential therapeutic for certain complement-mediated diseases. / Zhang, Lingjun; Armour, Kathryn; Chen, Jin Y. et al.
In: Molecular immunology, Vol. 170, 06.2024, p. 19-25.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaper › Journal article › Research › peer-review

TY - JOUR

T1 - Humanization of a mouse anti-human complement C6 monoclonal antibody as a potential therapeutic for certain complement-mediated diseases

AU - Zhang, Lingjun

AU - Armour, Kathryn

AU - Chen, Jin Y.

AU - Mylona, Agathi

AU - Yang, Maojing

AU - Andersen, Gregers R.

AU - Maciejewki, Jaroslaw P.

AU - Bakrania, Preeti

AU - Lin, Feng

PY - 2024/6

Y1 - 2024/6

N2 - The assembly of tissue-damaging membrane attack complexes (MACs; C5b–9) is a major mechanism by which excessive complement activation causes diseases. We previously developed a mouse anti-human C6 monoclonal antibody (mAb) 1C9 that selectively inhibits the assembly of MACs in human and non-human primates. In this project, we found that 1C9 also cross-reacted with rat and guinea pig C6, and determined its binding domains on C6 using different truncated C6 proteins. We then humanized the anti-C6 mAb by molecular modeling and complementarity-determining region grafting. After screening a library of 276 humanized variants with different combinations of humanized light and heavy chains in biophysical assays, we identified clone 3713 with the best developability profile, and an increased affinity against C6 when compared with the parental 1C9 mAb. This humanized 3713 mAb inhibited human, monkey, and rat complement-mediated hemolysis in vitro, and more importantly, it significantly reduced complement-mediated hemolysis in vivo in rats. These results demonstrated the successful humanization of the anti-C6 mAb and suggested that the humanized 3713 mAb could be further developed as a new therapeutic that selectively targets MAC for certain complement-mediated pathological conditions.

AB - The assembly of tissue-damaging membrane attack complexes (MACs; C5b–9) is a major mechanism by which excessive complement activation causes diseases. We previously developed a mouse anti-human C6 monoclonal antibody (mAb) 1C9 that selectively inhibits the assembly of MACs in human and non-human primates. In this project, we found that 1C9 also cross-reacted with rat and guinea pig C6, and determined its binding domains on C6 using different truncated C6 proteins. We then humanized the anti-C6 mAb by molecular modeling and complementarity-determining region grafting. After screening a library of 276 humanized variants with different combinations of humanized light and heavy chains in biophysical assays, we identified clone 3713 with the best developability profile, and an increased affinity against C6 when compared with the parental 1C9 mAb. This humanized 3713 mAb inhibited human, monkey, and rat complement-mediated hemolysis in vitro, and more importantly, it significantly reduced complement-mediated hemolysis in vivo in rats. These results demonstrated the successful humanization of the anti-C6 mAb and suggested that the humanized 3713 mAb could be further developed as a new therapeutic that selectively targets MAC for certain complement-mediated pathological conditions.

KW - Antibody

KW - C6

KW - Complement

KW - Humanization

KW - Guinea Pigs

KW - Humans

KW - Rats

KW - Cross Reactions/immunology

KW - Antibodies, Monoclonal, Humanized/immunology

KW - Complement Activation/immunology

KW - Animals

KW - Hemolysis/drug effects

KW - Antibodies, Monoclonal/immunology

KW - Complement Membrane Attack Complex/immunology

KW - Mice

KW - Complement C6/immunology

UR - http://www.scopus.com/inward/record.url?scp=85189797325&partnerID=8YFLogxK

U2 - 10.1016/j.molimm.2024.03.010

DO - 10.1016/j.molimm.2024.03.010

M3 - Journal article

C2 - 38598870

AN - SCOPUS:85189797325

SN - 0161-5890

VL - 170

SP - 19

EP - 25

JO - Molecular immunology

JF - Molecular immunology

ER -

Humanization of a mouse anti-human complement C6 monoclonal antibody as a potential therapeutic for certain complement-mediated diseases

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Keywords

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