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R&D Systems Biotinylated Proteins

R&D Systems Biotinylated Proteins Video

This video describes how R&D Systems amine-labeled and Avi-tag biotinylated recombinant proteins are produced and the advantages associated with either chemical or enzymatic biotinylation.

Advantages of Avi-Tag Biotinylated Proteins

Consistent, highly specific labeling. A single biotin molecule is enzymatically added to a lysine residue in the Avi-tag by BirA biotin ligase, resulting in the generation of a homogeneous product.

Uniform orientation of the protein. When bound to a streptavidin-coated surface, the orientation of the Avi-tag biotinylated protein will be uniform due to the precise control over biotinylation.

Similar bioactivity as the unlabeled protein. All R&D Systems biotinylated proteins are rigorously tested to ensure that they exhibit the same level of bioactivity as the unlabeled protein.

Lot-to-lot consistency. Each new lot is tested side-by-side with previous lots and with a master lot, so you don’t have to worry whether your results will be reproducible over time. 

Advantages of Amine-Labeled Biotinylated Proteins

• High signal strength. Chemical biotinylation labels the protein on amine groups found in lysine residues throughout the protein and at the N-terminus, resulting in multiple biotins being incorporated per protein.

Similar bioactivity as the unlabeled protein. All R&D Systems biotinylated proteins are rigorously tested to ensure that they exhibit the same level of bioactivity as the unlabeled protein.

Lot-to-lot consistency. Each new lot is tested side-by-side with previous lots and with a master lot, so you don’t have to worry whether your results will be reproducible over time. 

R&D Systems Biotinylated Proteins Exhibit the Same Levels of Bioactivity as the Equivalent Unlabeled Proteins

R&D Systems Avi-tag Biotinylated Recombinant Human PD-L1 has equivalent bioactivity as unlabeled Recombinant Human PD-L1

Unlabeled and Avi-tag Biotinylated Recombinant Human PD-L1/B7-H1 Display Comparable Bioactivity. Human T lymphocytes were treated with the indicated concentrations of either unlabeled Recombinant Human PD-L1/B7-1 Fc Chimera (R&D Systems, Catalog # 156-B7; green line) or Avi-tag Biotinylated Recombinant Human PD-L1/B7-H1 Fc Chimera (R&D Systems, Catalog # AVI156; orange line). IL-2 secretion was measured in cell culture supernatants using the Human IL-2 Quantikine® ELISA Kit (R&D Systems, Catalog # D2050). The similarity in the activities of the two proteins highlights that the Avi-tag biotinylated protein is fully functional.

R&D Systems Biotinylated Proteins Are Rigorously Tested to Ensure Lot-to-lot Consistency

R&D Systems Avi-tag Biotinylated Recombinant Human PD-L1/B7-H1 Displays High Lot-to-Lot Consistency. Three independent lots of Avi-tag Biotinylated Recombinant Human PD-L1/B7-H1 (R&D Systems, Catalog # AVI156) were tested for their ability to bind to Recombinant Human PD-1 (R&D Systems, Catalog # 1086-PD), which was coated at 1 ug/mL. Avi-tag Biotinylated Recombinant Human PD-L1/B7-H1 bound with an ED50 of 8-48 ng/mL. Each trace shown on the graph represents data obtained from Avi-tag Biotinylated Recombinant Human PD-L1/B7-H1 from a different manufacturing run to show the lot-to-lot consistency of the proteins.

Analysis of the lot-to-lot consistency of R&D Systems Avi-tag Biotinylated Recombinant Human PD-L1 Protein

Analysis of the Binding Properties of R&D Systems Avi-tag Biotinylated Proteins

Surface plasmon resonance data showing the affinity measurements and binding kinetics between the PD-L1 and PD-1 proteins.

Affinity Measurements and Binding Kinetics of the PD-1:PD-L1 Interaction by Surface Plasmon Resonance. Sensorgram data of captured Avi-tag Biotinylated Recombinant Human PD-L1 His tag (R&D Systems, Catalog # AVI9049) binding to Recombinant Human PD-1 His tag (R&D Systems, Catalog # 8986-PD). The corresponding overlaid kinetic fits with the residual plot shown below. The concentration of Recombinant Human PD-1 His-tag ranged from 3.2 nM to 13.2 μM. The corresponding steady state affinity fit is shown below. The experiment was performed on a BiacoreT200, GE Healthcare.

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