膜杰作
Star Staining
分子别名（Synonym）
HLA-A*02:01 & B2M & NY-ESO-1
表达区间及表达系统（Source）
Biotinylated Human HLA-A*02:01&B2M&NY-ESO-1 (SLLMWITQV) Complex Protein (HL1-H82E4) is expressed from human 293 cells (HEK293). It contains AA Gly 25 - Ile 308 (HLA-A*02:01) & Ile 21 - Met 119 (B2M) & SLLMWITQV peptide (Accession # AAA59606.1 (HLA-A*02:01) & P61769 (B2M) & SLLMWITQV).
Predicted N-terminus: Gly 25 & Ser
Request for sequence
蛋白结构（Molecular Characterization）
Biotinylated Human HLA-A*02:01&B2M&NY-ESO-1 (SLLMWITQV) Complex Protein is produced by co-expression of HLA and B2M loaded with NY-ESO-1 peptide.
This protein carries a polyhistidine tag at the C-terminus, followed by an Avi tag (Avitag™).
The protein has a calculated MW of 36.3 kDa and 13.8 kDa. The protein migrates as 42-45 kDa and 14 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.
标记（Labeling）
Biotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.
蛋白标记度（Protein Ratio）
Passed as determined by the HABA assay / binding ELISA.
内毒素（Endotoxin）
Less than 1.0 EU per μg by the LAL method.
纯度（Purity）
>95% as determined by SDS-PAGE.
>90% as determined by SEC-MALS.
制剂（Formulation）
Lyophilized from 0.22 μm filtered solution in PBS, pH7.4 with trehalose as protectant.
Contact us for customized product form or formulation.
重构方法（Reconstitution）
Please see Certificate of Analysis for specific instructions.
For best performance, we strongly recommend you to follow the reconstitution protocol provided in the CoA.
存储（Storage）
For long term storage, the product should be stored at lyophilized state at -20°C or lower.
Please avoid repeated freeze-thaw cycles.
This product is stable after storage at:
- -20°C to -70°C for 12 months in lyophilized state;
- -70°C for 3 months under sterile conditions after reconstitution.
电泳（SDS-PAGE）
Biotinylated Human HLA-A*02:01&B2M&NY-ESO-1 (SLLMWITQV) Complex Protein on SDS-PAGE under reducing (R) condition. The gel was stained with Coomassie Blue. The purity of the protein is greater than 95%.
SEC-MALS
The purity of Biotinylated Human HLA-A*02:01&B2M&NY-ESO-1 (SLLMWITQV) Complex Protein (Cat. No. HL1-H82E4) is more than 90% and the molecular weight of this protein is around 44-60 kDa verified by SEC-MALS.
Report
活性（Bioactivity）-ELISA
Immobilized Biotinylated Human HLA-A*02:01&B2M&NY-ESO-1 (SLLMWITQV) Complex Protein (Cat. No. HL1-H82E4) at 1 μg/mL (100 μL/well) on streptavidin (Cat. No. STN-N5116) precoated (0.5 μg/well) plate can bind Anti-NY-ESO-1 Antibody, Human IgG1 with a linear range of 0.2-16 ng/mL (QC tested).
Protocol
Immobilized Biotinylated Human HLA-A*02:01&B2M&NY-ESO-1 (SLLMWITQV) Complex Protein (Cat. No. HL1-H82E4) at 1 μg/mL (100 μL/well) on streptavidin (Cat. No. STN-N5116) precoated (0.5 μg/well) plate can bind Anti-HLA class I Antibody, Human IgG1 (W6/32) with a linear range of 0.1-2 ng/mL (Routinely tested).
Protocol
活性（Bioactivity）-SPR
Anti-NY-ESO-1 antibody captured on CM5 chip via Anti-human IgG Fc antibodies surface can bind Biotinylated Human HLA-A*02:01&B2M&NY-ESO-1 (SLLMWITQV) Complex Protein (Cat. No. HL1-H82E4) with an affinity constant of 8.36 nM as determined in a SPR assay (Biacore 8K) (QC tested).
Protocol
背景（Background）
NY-ESO-1, which is also well-known as New York esophageal squamous cell carcinoma 1, is an efficient target for cancer immunotherapy. This antigen is a member of cancer-testis antigens (CTAs) and is highly expressed in various cancers, including melanoma, ovarian, cervical cancer, etc. Adoptive T cell therapy with HLA-A2 restricted NY-ESO-1 transduced CD8+ T cells has improved the clinical response rates and overall survival of treatment-refractory melanoma patients. The Human HLA-A*0201 NY-ESO-1 (SLLMWITQV) complex protein is a complex of HLA-A*0201 of the MHC Class I, B2M and SLLMWITQV peptide of the NY-ESO-1.
