Avi-tag Biotinylated Recombinant Proteins

Avi-tag and chemically labeled biotinylated proteins

NEW Avi-tag Spike and ACE-2 Proteins for SARS-CoV-2 Research. Interested in more proteins for COVID-19 research? Visit our Recombinant Proteins for Coronavirus Research page.

R&D Systems offers a wide assortment of biotinylated proteins allowing researchers to take the guesswork out of protein biotinylation. In addition to amine-labeled biotinylated proteins, we have a growing list of Avi-tag biotinylated proteins. Each biotinylated protein is held to the same industry-leading quality standards as their unlabeled counterpart while also limiting batch-to-batch variability. Also, the R&D Systems Custom Protein Development Team can work with you to create a customized protein solution to meet your specific research needs.

Advantages of Avi-tag Biotinylated Recombinant Proteins

  • 1. Consistent labeling. Biotinylation only occurs on the single lysine residue in the Avi-tag.
  • 2. Uniform orientation of protein. When bound to a streptavidin-coated surface, the avitagged protein orientation will be uniform due to the precise control over biotinylation.
  • 3. Equivalent bioactivity. Because biotinylation is restricted to the Avi-tag, the rest of the protein is unchanged so there is no interference in the protein’s bioactivity.

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Avi-tag biotinylated recombinant protein binding to streptavidin-coated surface

Selected Avi-tag Proteins

For a complete up-to-date product range of all Avi-tag proteins

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5T4 CD28 EGFR IL-7R alpha/CD127 PDGF R alpha VEGF 165
ACE-2 CD30/TNFRSF8 EpCAM/TROP1 IL-12 R beta 1 PSMA/FOLH1/ NAALADase I VEGFR2/KDR/ Flk-1
B7-1/CD80 CD38 ErbB2/Her2 LAG-3 ROR1 VEGFR3/Flt-4
B7-2/CD86 CD40/TNFRSF5 Fc gamma RIIB/CD32b LAIR1 Siglec-2/CD22 VISTA/B7-H5/PD-1H
B7-H2 CD47 Fc gamma RIIIA LAIR2 Siglec-3/CD33 VSIG3
B7-H3 CD155/PVR GITR/TNFRSF18 Lymphotoxin alpha1/beta2 SIRP alpha/CD172a VSIG4
B7-H4 CD200 Glypican-3 M-CSF R/CD115 SARS-CoV-2 Spike VSTM2B
BCMA/TNFRSF17 CEACAM-5/ CD66e ICAM-1/CD54 MICA TIM-1/KIM-1/ HAVCR  
BTN1A1/Butyrophilin CTLA-4 IgG1 MSPR/Ron TIM-3  
BTN3A2 CX3CL1/ Fractalkine IL-2R beta PD-1 TSLPR  
CD19 CRACC/SLAMF7 IL-4R alpha PD-L1/B7-H1 uPAR  
CD25/IL-2R DNAM-1/CD226 IL-6R alpha PD-L2/B7-DC ULBP-2  

Amine-labeled and Sugar-labeled Biotinylated Proteins

We use R&D Systems best-in-class, highest quality recombinant proteins for biotinylation in an optimized process ensuring high labeling efficiency and lot-to-lot consistency. Proteins typically average multiple biotins per molecule making them an excellent choice when detection level is critical.

Custom Protein Libraries

Amine-labeled and Sugar-labeled Biotinylated Proteins

ACE-2 BMP-6 Ephrin-A3 IL-2 S5a/Angiocidin Viral B18R
Angiopoietin-1 BMP-7 Ephrin-A4 IL-6 SIRP alpha Viral B8R
Angiopoietin-2 AGE-BSA Ephrin-A5 ISG15 SARS-CoV-2 Spike VISTA
Angiopoietin-like 3, N-term BTN1A1 Ephrin-B1 LAG-3 SUMO2 VSIG3
Angiopoietin-like 4 BTN2A2 Ephrin-B2 LIF SUMO3 VSIG4
Angiopoietin-like 4, N-term BTN3A2 Ephrin-B3 NEDD8 TNF-alpha VSIG8
Angiopoietin-like 7 BTNL8 FAT10 PD-1 Ubiquitin Wnt-3a
Ataxin UIM CD155/PVR Frizzled-8  PD-L1 Poly-Ubiquitin Wnt-5a
B7-H3 CD277/BTN3A1 GABARAP PD-L2 Tetra-Ubiquitin  
B7-H4 Dkk-1 ICOS PDGF-BB TUBE1  
B7-H6 Ephrin-A1 IL-17A R-Spondin-1 TUBE2  
B7-H7 Ephrin-A2 IL-17A/F Heterodimer RAP80 UIM VEGF  

Biotinylated protein features:

Equivalent Bioactivity

Consistent bioactivity between biotinylated and non-biotinylated proteins

Both Biotinylated Recombinant Human PD-L1/B7-H1 Fc Avi-tag and unlabeled Recombinant Human PD-L1/B7-H1 inhibit anti-CD3 antibody-induced IL-2 secretion in human T lymphocytes
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Both Biotinylated Recombinant Human VEGF 165 and unlabeled Recombinant Human VEGF 165 stimulate HUVEC human umbilical vein endothelial cell proliferation
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Both Biotinylated Recombinant Human TNF-alpha and unlabeled Recombinant Human TNF-alpha promotes cytotoxicity in L‑929 mouse fibroblast cells in the presence of the metabolic inhibitor actinomycin D
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Both Biotinylated Recombinant Human PD-L1/B7-H1 Fc Avi-tag and unlabeled Recombinant Human PD-L1/B7-H1 inhibit anti-CD3 antibody-induced IL-2 secretion in human T lymphocytes. The similarity in activity highlights that the biotinylated Avi-tag protein is fully functional. Both Biotinylated Recombinant Human VEGF 165 (Catalog # BT293) and unlabeled Recombinant Human VEGF 165 (Catalog # 293-VE) stimulate HUVEC human umbilical vein endothelial cell proliferation. The ED50 for this effect is 1-6 ng/mL. The similarity in activity highlights that the biotinylated protein is fully functional. Both Biotinylated Recombinant Human TNF-alpha (Catalog # BT210) and unlabeled Recombinant Human TNF-alpha (Catalog # 210-TA) promotes cytotoxicity in L‑929 mouse fibroblast cells in the presence of the metabolic inhibitor actinomycin D. The ED50 for this effect is 25-100 pg/mL. The similarity in activity highlights that the biotinylated protein is fully functional.

Binding activity

When 500 ng/mL of Biotinylated Recombinant Human PD-1 His-tag Avi-tag protein is immobilized onto Streptavidin coated plate, it binds to Recombinant Human PD-L1/B7-H1 Fc Chimera with an ED50 of 1.2-7.2 ng/mL
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When Recombinant Human APRIL/TNFSF13 is immobilized at 0.1 µg/mL, Recombinant Human BCMA/TNFRSF17 Fc Chimera Avi-tag binds with an ED50 of 0.3-1.8 ng/m
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When Recombinant Human VSIG3 Fc Chimera is coated at 5 μg/mL, Biotinylated Recombinant Human VISTA/B7-H5/PD-1H Fc Chimera Avi-tag binds with an ED50 of 0.5-3 μg/mL.
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When 500 ng/mL of Biotinylated Recombinant Human PD-1 His-tag Avi-tag protein is immobilized onto Streptavidin coated plate (Catalog # CP004), it binds to Recombinant Human PD-L1/B7-H1 Fc Chimera (Catalog # 156-B7) with an ED50 of 1.2-7.2 ng/mL. When Recombinant Human APRIL/TNFSF13 (Catalog # 5860-AP) is immobilized at 0.1 µg/mL, Recombinant Human BCMA/TNFRSF17 Fc Chimera Avi-tag (Catalog # AVI193) binds with an ED50 of 0.3-1.8 ng/mL. When Recombinant Human VSIG3 Fc Chimera (Catalog # 9229-VS) is coated at 5 μg/mL, Biotinylated Recombinant Human VISTA/B7-H5/PD-1H Fc Chimera Avi-tag (Catalog # AVI7126) binds with an ED50 of 0.5-3 μg/mL.

Lot-to-lot consistency

When Recombinant Human PD-1 (Catalog #1086-PD) is coated at1 μg/mL, Biotinylated Recombinant Human PD-L1/B7-H1 Avi-tag (Catalog # AVI156) binds with an ED50 of 8-48 ng/mL.
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Three independent lots of B7-H1/PD-L1 have near identical bioactivity. When Recombinant Human PD-1 (Catalog #1086-PD) is coated at1 μg/mL, Biotinylated Recombinant Human PD-L1/B7-H1 Avi-tag (Catalog # AVI156) binds with an ED50 of 8-48 ng/mL.

Surface Plasmon Resonance (SPR)

BCD155/PVR-Fc Avi-tag protein
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Immobilized CD155/PVR-Fc Avi-tag (Catalog # AVI9174) bound to streptavidin CM5 chip binds to TIGIT-Fc (Catalog # 7898-TGB) with an affinity constant of 25.4 nM. Experiment performed on Biacore T200, GE Healthcare. For more information on using Avi-tag proteins in SPR, download the application note.

About Biotinylated Proteins

Biotinylated proteins are powerful tools used in a wide variety of assay formats, including immunoprecipitation, flow cytometry, immunoassays, biopanning, and surface plasmon resonance. Historically, individual labs have made their own biotinylated proteins in small batches. However, often times researchers encounter protein-specific technical challenges and high variability leading to inconsistent results.

What is Avi-tag?

Avi-tag Biotinylated Proteins are proteins that feature biotinylation at a single site contained within the Avi-tag, a unique 15 amino acid peptide. The DNA sequence coding for the Avi-tag is inserted during cloning and contained in the final expression vector construct. Biotinylation of the Avi-tag is enzymatically added by the E. coli biotin ligase BirA. This results in a single biotin molecule covalently attached to the lysine residue within the Avi-tag. The Avi-tag sequence is Gly-Leu-Asn-Asp-Ile-Phe-Glu-Ala-Gln-Lys-Ile-Glu-Trp-His-Glu.

Other biotinylated recombinant protein resources:

All proteins from R&D Systems

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