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Biotinylated Protein

INTRODUCTION

The interaction between biotin and avidin/streptavidin (SA) is incredibly strong and specific, making it an excellent tool in protein biochemistry. However, generation of high quality biotinylated proteins presents a major challenge. Researchers often run into technical issues including insufficient labeling, high batch-to-batch variation, and unpredictable performance. In addition, the choice of biotinylation methods also significantly affects the ultimate outcome. Each target protein (assay) has its own unique biochemical characteristics, and therefore requires customized solution.

These confounding factors make it difficult for individual labs to make and use biotin conjugates, while at the same time few products are available commercially. To fill in this gap, ACROBiosystems has developed an exclusive collection of pre-labeled and experimentally validated biotinylated recombinant proteins. The Mabsol® biotinylated protein collection includes more than a hundred commonly studied drug targets and biomarker proteins.

We use high quality recombinant proteins that are closest to natural conformation and modification as starting materials. We thoroughly go through every option to achieve the highest bioactivity and detection sensitivity. We also apply the most stringent quality control measurement possible to ensure minimal batch-to-batch variations.

Mabsol® biotinylated protein collection includes two unique and complimentary product series, the PrecisionAvi series built upon the AvitagTM technology (See full list) , and the UltraLys series produced using the in-house developed chemical labeling method (See full list). These products are made with every attention to details.

PRODUCTS

PrecisionAvi Series

An exclusive collection of ready-to-use AvitagTM biotinylated proteins

The products in this series are exclusively produced using the AvitagTM technology. Briefly, a unique 15 amino acid peptide, the Avi tag, is introduced into the recombinant protein during expression vector construction. The single lysine residue in the Avi tag is enzymatically biotinylated by the E. Coli biotin ligase BirA.

This single-point enzymatic labeling technique brings many advantages for commonly used binding assays.

  • The biotinylation only happens on the lysine residue of Avi tag.

  • NO interference with the target protein’s natural binding activities.

  • The protein orientation is uniform when immobilized on an avidin-coated surface.


Commonly searched products


Pre-order available for the following items: In additional to regular catalog products, we will also be accepting pre-orders for the following molecules, which are currently underdevelopment. Please contact us for technical specifics and price.


BACE CD147 CD319 CD22/SIGLEC2 CRP/C-Reactive CSF1R ErbB3 IL-5 IL-6 IL-1a IL-10 IL2R IL5R INSR LDLR SIRPA/CD172a TRAILR1 TRALR4 VEGFR1 TIM3/HAVCR2/KIM3

UltraLys Series

A unique series of chemically labeled biotinylated proteins with ultra sensitivity

The products in this series are produced using our in-house developed chemical labeling approach. The primary amines in the side chains of lysine residues and the N-terminus of protein are conjugated with biotins, usually resulting in multiple biotin attachment on a single protein molecule.

  • Higher detection sensitivity

  • In-house developed chemical labeling approach


Commonly searched products:

APPLICATIONS

DETECTION

ELISA

ELISA
Biotinylated proteins can be used in ELISA as two-way antibodies for both capture and detection with high specificity and detection sensitivity.

FACS Sorting

FACS
Biotinylated proteins can be used along fluorophore-tagged SA to detect/isolate cells expressing particular surface markers.

IMMOBILIZATION

Biopanning

Biopanning
Biotinylated proteins can be used with SA-coated magnetic beads/surface in biopanning with higher coating density and uniformed antigen presentation.

SPR

SPR
Biotinylated proteins can be used along with Biacore Sensor Chip SA for SPR analysis with low baseline drift and noise, and low activity loss during surface regeneration cycle.

Enrichment

Enrichment
Biotinylated proteins can be used to isolate antibodies from plasma or other biological fluid for subsequent analyses with high sensitivity, and the sample can be easily analyzed in quantitative mass spectrometry.
KEY FEATURES

Closest to Natural Conformation and Modification

The production of our recombinant proteins including the biotinylated proteins is carried out using our proprietary HEKMax® expression platform. As expression hosts, the human HEK293 cells have a variety of advantages compared to other cell types as summarized in the table below.


Expression Systems
E.Coli
Insect Cell
Plant Cell
CHO Cell
Human Cell
Folding
+
++
+++
++++
+++++
Phosphorylation
N/A
++
+++
++++
+++++
Proteolytic Processing
N/A
++
+++
++++
+++++
Glycosylation
N/A
Poor
Poor
Non-Human Like
Human Authentic

High Bioactivity & Detection Sensitivity

The bioactivity of biotinylated proteins is determined both by the structure of the protein itself, and by the way how biotinylation is performed. For every single protein, we test multiple options of tags and biotinylation methods, and evaluate the products in a variety of binding assays. Only those with the best performance are selected for production. Figure 1 is an example of our internal evaluation experiments:


Fig. 1 Binding activity of different forms of biotinylated PD1 evaluated in a functional ELISA against rhPD-L1 (Cat. No. PD1-H5258).


Our biotinylated proteins demonstrate high bioactivity and superb detection sensitivity. In the following Fig. 2, Biotinylated FcRn / FCGRT & B2M (Cat. No. FCM-H82W4) coupled to SA coated sensor chip can bind Herceptin with an affinity constant of 2.24 μM as determined in a SPR assay.


Fig. 2 Biotinylated Human FcRn / FCGRT & B2M, Avi Tag (Avitag™) (Cat. No. FCM-H82W4) coupled to SA coated sensor chip can bind Herceptin with an affinity constant of 2.24 μM as determined in a SPR assay (Biacore 2000). The data is generally provided by Biaffin GmbH &Co KG, Germany.

Fig. 3 Immobilized Human BAFF, Fc Tag (Cat. No. BAF-H4268) at 5 μg/mL (100 μL/well) can bind Biotinylated Human BCMA, Fc Tag (Cat. No. BC7-H82F0) with a linear range of 0.12-1.95 ng/mL (QC tested).

Low Batch-to-Batch Variation

We routinely apply rigorous quality control measures to ensure consistent performance of our products. Newly produced products are subjected to side-by-side comparison with our internal standard in a variety of assays. Only those within an acceptable margin of difference are allowed to be released. As demonstrated in Fig. 4, three different batches of biotinylated hTNF-alpha (Cat. No. TNA-H82E3) are tested and compared using a standard ELISA analysis against Adalimumab and 0.5µg/well Adalimumab was used to coat the plate, the result showed that the batch variation among the tested samples is negligible.

Fig. 4 In the above ELISA analysis, three different lots of biotinylated hTNF-alpha (Cat. No. TNA-H82E3) were used detect immobilized Adalimumab (5ug/ml). The result showed that the batch variation among the tested samples is negligible.


LATEST CITATIONS

Integration of a CD19 CAR into the TCR Alpha Chain Locus Streamlines Production of Allogeneic Gene-edited CAR T Cells

  • Authors: MacLeod DT et al.

  • Journal: Mol Ther 2017

  • Product: biotinylated Human CD19(Cat. No. CD9-H8259

  • Application: Flow Cytometry

  • Request for Full-text

Quantification of Active Infliximab in Human Serum with Liquid Chromatography-Tandem Mass Spectrometry Using a Tumor Necrosis Factor Alpha -based Pre-analytical Sample Purification and a Stable Isotopic Labeled Infliximab Bio-similar as Internal Standard: A Target-based, Sensitive and Cost-effective Method

  • Authors: El Amrani M et al.

  • Journal: J Chromatogr A 2016

  • Product: biotinylated human recombinant TNF-alpha(Cat. No. TNA-H8211

  • Application: ELISA

  • Request for Full-text

Antibody-drug Conjugate Having Improved Stability and Use Thereof

  • Authors: Kim Young Min et al.

  • Journal: United States Patent Application 20160136300 2016

  • Product: ErbB2(Cat. No. HE2-H822R

  • Application: ELISA

  • Request for Full-text

A Simplified Procedure for Antibody Engineering by Yeast Surface Display: Coupling Display Levels and Target Binding by Ribosomal Skipping

Microsphere-based Immunoassay Integrated with a Microfluidic Network to Perform Logic Operations

  • Authors: Pooja Sabhachandani et al.

  • Journal: Microchimica Acta 2015

  • Product: biotinylated human TNF-α(Cat. No. TNA-H8211

  • Application: Microsphere-based assay

  • Request for Full-text

The Shark Strikes Twice: Hypervariable Loop 2 of Shark IgNAR Antibody Variable Domains and Its Potential to Function as an Autonomous Paratope

  • Authors: Zielonka S et al.

  • Journal: Marine Biotechnology 2015

  • Product: CD3 epsilon(Cat. No. CDE-H8223

  • Application: FACS sorting

  • Request for Full-text

Novel Cell Line Screening Method

  • Authors: YU et al.

  • Journal: United States Patent Application 20140155285 2014

  • Product: biotinylated TNFα(Cat. No. TNA-H8211

  • Application: Flow Cytometry

  • Request for Full-text

LIMITED USE LICENSE
The Biotin AviTag technology is covered by U.S. Pat. No: 5,874,239 and includes any and all materials, methods, kits and related derivatives claimed by this patent. The purchase of the ACROBiosystems’s AvitagTM proteins confers to the purchaser the limited right to use the AvitagTM technology for non-commercial, or research use, or for purposes of evaluating the AvitagTM technology.

Commercial use of the AvitagTM technology to manufacture a commercial product, or use of the AvitagTM technology to facilitate or advance research which will be applied to the development of a commercial product requires a license from Avidity, LLC. Examples of Commercial use include, but are not limited to biosensors, diagnostics, therapeutic applications, proximity assays, and drug screening assays.

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