Biotinylated Human HLA-A*02:01&B2M&MAGE-A4 (KVLEHVVRV) Complex Protein (Monomer, MALS verified)

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INTRODUCTION

QC & Assay Protocol

Citation & Background

Source
Biotinylated Human HLA-A*02:01&B2M&MAGE-A4 (KVLEHVVRV) Complex Protein(HLP-H82E5) is expressed from human 293 cells (HEK293). It contains AA Gly 25 - Ile 308 (HLA-A*02:01) & Ile 21 - Met 119 (B2M) & KVLEHVVRV peptide (Accession # AAA59606.1 (HLA-A*02:01) & P61769-1 (B2M) & KVLEHVVRV).
Predicted N-terminus: Gly 25 & Lys
Molecular Characterization
Biotinylated Human HLA-A*02:01&B2M&MAGE-A4 (KVLEHVVRV) Complex Protein is produced by co-expression of HLA and B2M loaded with MAGE-A4 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 15 kDa under reducing (R) condition (SDS-PAGE) due to glycosylation.
Labeling
Biotinylation of this product is performed using Avitag™ technology. Briefly, the single lysine residue in the Avitag is enzymatically labeled with biotin.
Protein Ratio
Passed as determined by the HABA assay / binding ELISA.
Endotoxin
Less than 1.0 EU per μg by the LAL method.

Purity
>90% as determined by SDS-PAGE.
>95% as determined by SEC-MALS.
Formulation
Lyophilized from 0.22 μm filtered solution in PBS, pH7.4 with trehalose as protectant.
Contact us for customized product form or formulation.
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.
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:
1. -20°C to -70°C for 12 months in lyophilized state;
2. -70°C for 3 months under sterile conditions after reconstitution.

SDS-PAGE

Biotinylated Human HLA-A*02:01&B2M&MAGE-A4 (KVLEHVVRV) Complex Protein on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 90%.

SEC-MALS

HLA-A*0201 | B2M & MAGE-A4 (KVLEHVVRV) MALS images

The purity of Biotinylated Human HLA-A*02:01&B2M&MAGE-A4 (KVLEHVVRV) Complex Protein (Cat. No. HLP-H82E5) is more than 95% and the molecular weight of this protein is around 45-60 kDa verified by SEC-MALS.

Report

Background
The MAGE A4 antigen is a cancer-testis antigen and is expressed intracellularly in various solid tumor tissues, MAGE A4230-239 peptide (GVYDGREHTV) is a cytotoxic T lymphocyte (CTL) epitope presented by HLA-A2 The Human HLA-A*0201 MAGE-A4 (GVYDGREHTV) complex protein is a complex of HLA-A*0201 of the MHC Class I, B2M, and GVYDGREHTV peptide of the MAGE-A4.
Clinical and Translational Updates
Please contact us via TechSupport@acrobiosystems.com if you have any question on this product.