Anti-PD-L1 antibody, human IgG1, recombinant

Description

PD-L1 (programmed cell death 1 ligand 1) or CD274 is a 290-amino acid single pass type I transmembrane protein of the Ig (immunoglobulin) superfamily.  It contains one Ig V-like and one Ig C-like domains in the extracellular region, and a 30-amino acid cytoplasmic tail. It was originally cloned as a homolog of B7 (termed B7-H1). It is 20% and 15% identical to B7-1 (CD80) and B7-2 (CD86), respectively.  It was also cloned as a ligand (termed PDCD1L1 or PD-L1) that binds to PD-1 (CD279) and B7-1 (CD80), but not to other B7 family members, such as CTLA4 (CD152), CD28, or ICOS (CD278).  PD-L1 or CD274 is involved in the T cell costimulatory signal, essential for T-cell proliferation and production of IL10 and IFNγ. Interaction with PD-1, which has a cytoplasmic immunoreceptor tyrosine-based inhibitory motif (ITIM), inhibits T-cell proliferation and cytokine production.  PD-L1 is up-regulated on T- and B-cells, dendritic cells, keratinocytes and monocytes after LPS and IFNγ activation or by surface Ig cross-linking. It is expressed in many types of freshly isolated human cancers but not in most normal tissues.  Blockade of PD-L1 upregulates IL12 expression and enhances antitumor immunity in mice bearing ovarian tumors.  Therefore, PD-L1 has been proposed as a target for cancer immunotherapy through pre-activated T cells that could be enhanced by blockade of PD-L1-induced inhibitory signaling. 

 

 

Gene Symbol: PD-L1; CD274; B7-H1; B7H1;  PDL1;  PDCD1L1; PDCD1LG1

 

NCBI Gene ID: 29126 (human), 60533 (mouse)

 

Uniprot Entry: Q9NZQ7 (human), Q9EP73 (mouse)

 

Construct Details: The recombinant antibody is expressed as the combination of a heavy chain (HC) containing VH from anti-PD-L1 mAb and CH1-3 region of human IgG1 and a light chain (LC) encoding VL from anti-PD-L1 mAb and CL of human kappa light chain.  Exists as a disulfide linked dimer of the HC and LC hetero-dimer under non-reducing condition.

 

Source: Mammalian cells stably expressing anti-PD-L1 HC and LC and growing in chemical-defined media with no animal component or antibiotics.

 

M.W.:  Calculated molecular mass (kDa): 146 (49 kDa for a single HC, 24 kDa for a single LC); Estimated by SDS-PAGE under non-reducing condition(kDa): 150.

 

Purity: >95% judged by SDS-PAGE under reducing condition (see the gel image above)

 

Formulation: Supplied at 1.0 mg/ml in sterile PBS pH7.4 without any carrier protein or preservative (the concentration was determined by UV absorption with an A280nm of 1.0 for 0.73 mg/ml of IgG and verified by SDS-PAGE and Coomassie blue staining).

 

Endotoxin Level: <0.1 EU per 1 μg of purified recombinant protein determined by the LAL method

 

Biological Activity: Binds to human PD-L1 extracellular domain protein (SKU#FCL0784) and PD-L1-Fc  (SKU#FCL0781) with high affinity (K_D < 1 nM as determined by ELISA).  Cross-react with mouse and monkey PD-L1 with high affinity similar to human PD-L1's binding. Blocks PD-L1’s binding to its receptor PD-1 and B7-1/CD80 and neutralizes PD-L1's signaling activity. Enhances T-cell-mediated anti-tumor immunity.  Binds PD-L1 with the same binding specificity as anti-PD-L1 mouse IgG2a (MAB0784) and rabbit IgG (MAB0781).

 

 

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE IN HUMAN.

Restriction: This product is not transferable or re-sellable.  Customer obtain no right to transfer, assign, or sublicense its use rights, or to transfer, resell, package, or otherwise distribute the product, or to use the product for the benefit of any third party or for any commercial purpose.  Customer may only use the product in compliance with applicable local, state and federal laws, regulations and rules.  Customer may not directly or indirectly use the product or allow the transfer, transmission, export or re-export of all or any part of the product in violation of any export control law or regulation of the united states or any other relevant jurisdiction.  Your use of this product constitutes acceptance of the terms of this limited use agreement.  Please refer to our “terms & conditions” for details.  If you are not willing to accept the limitation of this agreement, G&P Biosciences will accept return of the product for a full/partial refund.

  

 

Technical Data 1:  Determination of Binding Affinity to Human PD-L1 by ELISA

Recombinant PD-L1 ECD protein (SKU#FCL0784) was immobilized at 150 ng/well on a 96-well NuncMaxisorp plate for an ELISA with serially diluted anti-PD-L1 monoclonal antibody (SKU#MAB0199). Anti-PD-1 (SKU#MA1732) monoclonal antibody was used as a negative control to demonstrate the specificity of PD-L1 binding under the experimental conditions.  The binding curve was fit and apparent K_D was calculated for anti-PD-L1 : PD-L1 binding to be 21.6 ± 1.1 pM.   

 

 

Technical Data 2: Cross-species Interactions with Human and Mouse PD-L1 by ELISA

Recombinant human PD-L1-Fc (hPD-L1, SKU#FCL0781), mouse PD-L1-Fc (mPD-L1, SKU#FCL1846), human PD-L2-Fc (hPD-L2, SKU#FCL0783), and mouse PD-L2-Fc (mPD-L2, SKU#FCL1818) proteins was immobilized at 100 ng/well on a 96-well NuncMaxisorp plate for ELISA with serially diluted, biotinylated anti-PD-L1 monoclonal antibody, human IgG1(SKU#MAB0199B). PD-L2 proteins were used as a control to demonstrate the specificity of PD-L1 binding under the experimental conditions. As shown in the figure below, anti-PD-L1 mAb binds specifically to human and mouse PD-L1-Fc (but not to PD-L2-Fc) equally well with an apparent K_D for human PD-L1 of 38.1 ± 2.4 pM and an apparent K_D for mouse PD-L1 of 38.6± 2.5 pM.

 

 

 

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE IN HUMAN.

 

 

The product is shipped at 4°C. Upon receipt, centrifuge the product briefly before opening the vial. It is recommended to store small aliquots at the temperature below –20°C for long-term storage and the product is stable for 3 months. The undiluted protein can be stored at 4°C for no more than 2 weeks.

 

 

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.

 

 

 

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE IN HUMAN.

 

 

1. Nat. Med. 5:1365-1369, 1999

2. J. Exp. Med. 192:1027-1034, 2000

3. Nature Med. 8: 793-800, 2002

4. Nature Med. 9: 562-567, 2003

 

 

 

Product datasheet (pdf) can be downloaded here: MAB0199-PDS.pdf

 

 

 

Additional supporting documents, including COA and MSDS are available upon request.

 

 

 

FOR RESEARCH USE ONLY. NOT FOR DIAGNOSTIC OR THERAPEUTIC USE IN HUMAN.

Restriction: This product is not transferable or re-sellable.  Customer obtain no right to transfer, assign, or sublicense its use rights, or to transfer, resell, package, or otherwise distribute the product, or to use the product for the benefit of any third party or for any commercial purpose.  Customer may only use the product in compliance with applicable local, state and federal laws, regulations and rules.  Customer may not directly or indirectly use the product or allow the transfer, transmission, export or re-export of all or any part of the product in violation of any export control law or regulation of the united states or any other relevant jurisdiction.  Your use of this product constitutes acceptance of the terms of this limited use agreement.  Please refer to our “terms & conditions” for details.  If you are not willing to accept the limitation of this agreement, G&P Biosciences will accept return of the product for a full/partial refund.