BTLA/HVEM Pathway: Emerging Checkpoint Biology Driving Next-Gen Immunotherapy

BTLA-HVEM Interaction Network
（https://doi.org/10.1016/j.ejmech.2024.116231）

The purity of Human LIGHT Protein, Fc Tag (Cat. No. LIT-H5269) is more than 95% and the molecular weight of this protein is around 180-200 kDa verified by SEC-MALS.

Immobilized Human BTLA (31-150), His Tag (Cat. No. BTA-H52H3) at 5 μg/mL (100 μL/well) can bind Human HVEM, Fc Tag (Cat. No. HVM-H5258) with a linear range of 0.005-0.313 μg/mL (QC tested).

Immobilized Human LIGHT, Mouse IgG2a Fc Tag, low endotoxin (Cat. No. LIT-H5256) at 2 μg/mL (100 μL/well) can bind Biotinylated Human HVEM, Fc, Avitag (Cat. No. HVM-H82F6) with a linear range of 0.6-39 ng/mL (QC tested).

Loaded Human BTLA (31-150), His Tag (Cat. No. BTA-H52H3) on NTA Biosensor, can bind Human HVEM, Mouse IgG2a Fc Tag, low endotoxin with an affinity constant of 20.8 nM as determined in BLI assay (ForteBio Octet Red96e) (Routinely tested).

Human LIGHT Protein, His Tag (Cat. No. LIT-H5242) induced cytotoxicity in HT-29 cells. The ED50 for this effect is 1.11-3.87 ng/mL (Routinely tested).

Expression analysis of human HVEM on Human HVEM (Luc) HEK293 Reporter Cell by FACS.Cell surface staining was performed on Human HVEM (Luc) HEK293 Reporter Cell (Cat. No. CHEK-ATF105) or negative control cell using PE-labeled anti-human HVEM antibody

Inhibition of human BTLA overexpressing on CHO cells induced reporter activity by anti-human BTLA antibody.This reporter cell was incubated with serial dilutions of antibodies in the presence of CHO/Human BTLA Stable Cell Line (Cat. No. SCCHO-ATP110). The EC50 of anti-human BTLA neutralizing antibody is approximately 0.212 μg/mL.

Passage stability analysis by Signaling Bioassay. The continuously growing Human HVEM (Luc) HEK293 Reporter Cell (Cat. No. CHEK-ATF105) was incubated with CHO/Human BTLA Stable Cell Line (Cat. No. SCCHO-ATP110). The human BTLA-overexpressing CHO cells stimulated response demonstrates passage stabilization (fold induction) across passage 7-25.

Passage stability analysis of receptor expression by FACS. Flow cytometry surface staining of human BTLA on CHO/Human BTLA Stable Cell Line (Cat. No. SCCHO-ATP110) demonstrates consistent mean fluorescent intensity across passage 3-20.