B-hPD-L1 plus/hHER2 MC38_Biocytogen Pharmaceuticals (Beijing) Co., Ltd._Biocytogen, drug development, antibody discovery

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B-hPD-L1 plus/hHER2 MC38

Common name	B-hPD-L1 plus/hHER2 MC38	Catalog number	321917
Aliases	CD274, B7-H, B7H1, PD-L1, PDCD1L1, PDCD1LG1, PDL1, hPD-L1, CD274 molecule,Programmed cell death 1 ligand 1; ERBB2, CD340, HER-2, HER-2/neu, HER2, MLN 19, NEU, NGL, TKR1, VSCN2, c-ERB-2, c-ERB2, erb-b2 receptor tyrosine kinase 2	Disease	Colon carcinoma
Organism	Mouse	Strain	C57BL/6
Tissue types	Colon	Tissue	Colon

Description

The mouse Pdl1 gene was replaced by human PD-L1 coding sequence in B-hPD-L1 plus/hHER2 MC38 cells. Human PD-L1 is highly expressed on the surface of B-hPD-L1 plus/hHER2 MC38 cells.

The mouse Erbb2 gene was replaced by human ERBB2 coding sequence in B-hPD-L1 plus/hHER2 MC38 cells. Human ERBB2 is highly expressed on the surface of B-hPD-L1 plus/hHER2 MC38 cells.

Application

B-hPD-L1 plus/hHER2 MC38 cells have the capability to establish tumors in vivo and can be used for efficacy studies.

Targeting strategy

Gene targeting strategy for B-hPD-L1 plus/hHER2 MC38 cells.
The exogenous promoter and human PD-L1 coding sequence was inserted to replace part of murine exon 3. The insertion disrupts the endogenous murine Pdl1 gene, resulting in a non-functional transcript.
The exogenous CAG promoter and human ERBB2 coding sequence was inserted to replace part of murine exon 2 and all of exons 3-7. The insertion disrupts the endogenous murine Erbb2 gene, resulting in a non-functional transcript.

Protein expression analysis

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PD-L1 and HER2 expression analysis in B-hPD-L1 plus/hHER2 MC38 cells by flow cytometry. Single cell suspensions from wild-type MC38 and B-hPD-L1 plus/hHER2 MC38 cultures were stained with species-specific anti-PD-L1 and anti-HER2 antibody. Human PD-L1 and HER2 were detected on the surface of B-hPD-L1 plus/hHER2 MC38 cells but not wild-type MC38 cells. The 3-B08 clone of B-hPD-L1 plus/hHER2 MC38 cells was used for in vivo tumor growth assays.

Tumor growth curve & Body weight changes

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Subcutaneous homograft tumor growth of B-hPD-L1 plus/hHER2 MC38 cells. B-hPD-L1 plus/hHER2 MC38 cells (5x10⁵) and wild-type MC38 cells (5x10⁵) were subcutaneously implanted into C57BL/6 mice (female, 9-week-old, n=6). Tumor volume and body weight were measured twice a week. (A) Average tumor volume ± SEM. (B) Body weight (Mean ± SEM). Volume was expressed in mm³ using the formula: V=0.5 X long diameter X short diameter². As shown in panel A, B-hPD-L1 plus/hHER2 MC38 cells were able to form tumors in vivo and can be used for efficacy studies.

Tumor growth curve of individual mice

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B-hPD-L1 plus/hHER2 MC38 MC38 tumor cells growth of individual mice. B-hPD-L1 plus/hHER2 MC38 cells (5x10⁵) and wild-type MC38 cells (5x10⁵) were subcutaneously implanted into C57BL/6 mice (female, 9-week-old, n=6). As shown in panel, B-hPD-L1 plus/hHER2 MC38 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.

Protein expression analysis of tumor cells

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B-hPD-L1 plus/hHER2 MC38 cells were subcutaneously transplanted into C57BL/6 mice (n=6), and on 39 days post inoculation, tumor cells were harvested and assessed for human PD-L1 and HER2 expression by flow cytometry. As shown, human PD-L1 and HER2 were highly expressed on the surface of tumor cells. Therefore, B-hPD-L1 plus/hHER2 MC38 cells can be used for in vivo efficacy studies of PD-L1 and HER2 therapeutics.

Tumor growth curve & Body weight changes

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Subcutaneous homograft tumor growth of B-hPD-L1 plus/hHER2 MC38 cells. B-hPD-L1 plus/hHER2 MC38 cells (2x10⁵, 5x10⁵) and wild-type MC38 cells (5x10⁵) were subcutaneously implanted into homozygous B-hHER2 mice (female, 5-week-old, n=6). Tumor volume and body weight were measured twice a week. (A) Average tumor volume ± SEM. (B) Body weight (Mean ± SEM). Volume was expressed in mm³ using the formula: V=0.5 X long diameter X short diameter². As shown in panel A, B-hPD-L1 plus/hHER2 MC38 cells were able to form tumors in vivo and can be used for efficacy studies.

Tumor growth curve of individual mice

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B-hPD-L1 plus/hHER2 MC38 MC38 tumor cells growth of individual mice. B-hPD-L1 plus/hHER2 MC38 cells (2x10⁵, 5x10⁵) and wild-type MC38 cells (5x10⁵) were subcutaneously implanted into homozygous B-hHER2 mice (female, 5-week-old, n=6). As shown in panel, B-hPD-L1 plus/hHER2 MC38 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.

Tumor volume and weight measurements

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Protein expression analysis of tumor cells

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B-hPD-L1 plus/hHER2 MC38 cells were subcutaneously transplanted into homozygous B-hHER2 mice (n=6), and on 30 days post inoculation, tumor cells were harvested and assessed for human PD-L1 and HER2 expression by flow cytometry. As shown, human PD-L1 and HER2 were highly expressed on the surface of tumor cells. Therefore, B-hPD-L1 plus/hHER2 MC38 cells can be used for in vivo efficacy studies of PD-L1 and HER2 therapeutics.