Basic Information
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Gene targeting strategy
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Gene targeting strategy for B-hCD40/hFcRn mice. Exons 2-7 of mouse Cd40 gene that encode the extracellular sequence were replaced by human CD40 exons 2-7 in B-hCD40/hFcRn mice. The human full-length FCGRT cDNA sequence was inserted in the Fcgrt exons 2-4 in B-hCD40/hFcRn mice.
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Protein expression analysis
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Species-specific CD40 protein expression analysis in wild-type and humanized B-hCD40/hFcRn mice. Splenocytes were isolated from wild-type C57BL/6 (+/+) and homozygous B-hCD40/hFcRn (H/H) mice and analyzed by flow cytometry using species-specific anti-CD40 antibodies. Murine CD40 protein was detected in wild-type mice, while human CD40 protein was detected in B-hCD40/hFcRn mice.
Species-specific FcRn protein expression analysis in wild-type and humanized B-hCD40/hFcRn mice. Liver tissue was isolated from wild-type C57BL/6 (+/+) and homozygous B-hCD40/hFcRn (H/H) mice and analyzed by western blot using species-specific anti-FcRn antibodies. Murine FcRn protein was detected in wild-type mice, while human FcRn protein was detected in B-hCD40/hFcRn mice.
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Determination of plasma concentrations of human IgG in different mice
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The study evaluated the pharmacokinetic characteristics of anti-human CD40 antibodies in B-hCD40/hFcRn mice. C57BL/6 mice, B-hFcRn mice, and B-hCD40/hFcRn mice were treated with YH015 or YH003 (anti-human CD40 antibodies) (n=6). Blood samples were collected at different time points for the pharmacokinetic (PK) assay. Compared to wild-type mice and B-hFcRn mice, B-hCD40/hFcRn mice can more accurately reflect the in vivo concentration changes of anti-human CD40 antibodies, the results showed that B-hCD40/hFcRn mice can be used to evaluate the pharmacokinetics of anti-human CD40 antibodies in vivo.
