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B-hCLDN18.2 A549
Common name
B-hCLDN18.2 A549 Catalog number    310791
Aliases OASFTA5, SFTPJ Disease  Carcinoma
Organism
Human Strain  --
Tissue types Lung Tissue  Lung

Description


The human CLDN18.2 coding sequence was inserted into the AAVS1 locus in A549 cells. Human CLDN18.2 is highly expressed on the surface of B-hCLDN18.2 A549 cells.


Application


B-hCLDN18.2 A549 cells have the capability to establish tumors in vivo and can be used for efficacy studies.


Targeting strategy


Gene targeting strategy for B-hCLDN18.2 A549 cells. The exogenous promoter and human CLDN18.2 coding sequence was inserted into the AAVS1 locus wild-type A549.


Protein expression analysis

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CLDN18.2 expression analysis in B-hCLDN18.2 A549 cells by flow cytometry. Single cell suspensions from wild-type A549 and B-hCLDN18.2 A549 cultures were stained with species-specific anti-CLDN18.2 antibody. Human CLDN18.2 was detected on the surface of B-hCLDN18.2 A549 cells but not wild-type A549 cells. The 1-A04 clone of B-hCLDN18.2 A549 cells was used for in vivo experiments.

Tumor growth curve & Body weight changes


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Subcutaneous heterograft tumor growth of B-hCLDN18.2 A549 cells. B-hCLDN18.2 A549 cells (1x107) and wild-type A549 cells (1x107) were subcutaneously implanted into B-NDG mice (female, n=5). 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-hCLDN18.2 A549 cells were able to establish tumors in vivo and can be used for efficacy studies.

Protein expression analysis in tumor cells

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B-hCLDN18.2 A549 cells were subcutaneously transplanted into B-NDG mice (n=5). At the end of the experiment, tumor cells were harvested and assessed for human CLDN18.2 expression by flow cytometry. As shown, human CLDN18.2 was highly expressed on the surface of tumor cells. Therefore, B-hCLDN18.2 A549 cells can be used for in vivo efficacy studies of novel CLDN18.2 therapeutics.


In vivo Efficacy of anti-hCLDN18.2 antibodies 

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Antitumor activity of anti-hCLDN18.2 antibodies in CB-17 SCID mice. (A) Anti hCLDN18.2 antibody slightly inhibited A549-hCLDN18.2 tumor growth in CB-17 SCID mice. B-CAG-hCLDN18.2 A549 cells were subcutaneously implanted into CB-17 SCID mice (female, 7 week-old, n=5). Mice were grouped when tumor volume reached approximately 90 mm3, at which time they were treated with anti-hCLDN18.2 antibody with different doses and schedules indicated in panel (B) Body weight changes during treatment. As shown in panel A, anti-hCLDN18.2 antibody was efficacious but mild, demonstrating that B-CAG-hCLDN18.2 A549 cells can be used to establish tumor model and provide a powerful preclinical model for in vivo evaluation of anti-hCLDN18.2 antibodies. Values are expressed as mean ± SEM.