Preclinical hCD3ε Model for Assessment of T-cell Engager Efficacy

Humanized CD3 Mouse Models

Design of the hCD3ε mouse

The humanized CD3ε mouse model was developed to express the human epitope of the CD3ε chain, which is recognized by T-cell engagers (clone SP34). The rest of the extracellular domain is murine to preserve the amino acids involved in the interaction with CD3γ and CD3δ.

Similarly, the transmembrane domains and the intracellular domains are murine to enable salt bridges’ interaction, interaction with the CD3ζ, and the signaling into mouse cells.

Applications in preclinical studies

The hCD3ε mouse enables the in vivo efficacy assessment of T-cell engagers in fully immunocompetent mice.


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hCD3ε features

  • Extracellular CD3ε N-terminal epitope is humanized
  • Conserved interaction with CD3γ, CD3δ and CD3ζ interactions
  • Fully functional mouse immune system

See also:

hCD3ε validation*

The model has been co-validated by our partners of leaders in immuno-oncology and immunotherapy.

A scientific poster was presented at SITC Cancer Immunotherapy CONNECT 2020. Request the poster

Double-positive cells are identified with similar levels from homozygous mice using clone SP34 against hCD3ε, and from wild-type using anti-mouse CD3ε, which binds to CD3ε, interacting with CD3γ or CD3δ chains

hCD3epsilon model validation 1

SP34 epitope of CD3ε is humanized. All other components of the CD3 complex are mouse.

(A) Diagram showing the co-expression of human CD3ε N-terminal epitope with mouse CD3 complex. (B) Splenocytes were isolated from wild-type and homozygous hCD3ε mice. Expression of CD3ε N-terminal human epitope recognized by the SP34.2, as well as expression of mouse CD3ε, was assessed using anti-human CD3 (clone SP342) and anti-mouse CD3 (clone 145-2C11), respectively, on single live splenocytes (n=3).

hCD3epsilon model validation 2

A T-cell engager targeting CD3 and a tumoral antigen, showed a dose-dependent anti-tumor effect in vivo

T-cell engagers efficacy assessment. (A) Experimental design for in vivo experiments. Tumor growth curves in (B) wild-type and (C) hCD3ε mice.

* For more validation data please contact us.


Further reading


Ready to be shipped to your lab

  • Cohorts available upon request
  • Studies can be carried out at your site or at your favorite CRO
  • SOPF certification and worldwide delivery by professional breeders
  • Models provided with FTO on patent-protected technologies used for model generation

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