Recombinant Human IL-2 (CD122-Directed) Protein, CF New

Catalog #: BT-002DBR Datasheet / COA / SDS

AI Modified Protein

Catalog #	Availability	Size / Price	Qty
BT-002DBR-010
BT-002DBR-050
BT-002DBR-100
BT-002DBR-500
BT-002DBR-01M

IL-2 (CD122-Directed) Provides Increased and Sustained CD4+/CD8+ T Cell Proliferation in Side-by-Side Testing with the Standard IL-2 Protein

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Summary Product Datasheets Carrier Free Data Images Reconstitution Calculator Background Related Research Areas

Recombinant Human IL-2 (CD122-Directed) Protein, CF Summary

Il-2 receptor engagement can impact the growth and phenotype of cultured immune cells such as T, NK, or Tumor Infiltrating T cells. By engaging the beta receptor (CD122) with higher affinity, continued proliferation of T cells has been observed. Utilizing generative AI to develop an optimized IL-2 cytokine with enhanced beta receptor (CD122) engagement offers superior performance while maintaining greater than 95% homology to its wild-type counterpart including:

Enhanced CD122 Binding: Increased affinity to the IL-2 beta receptor (CD122) with decreased affinity to IL-2 alpha receptor (CD25), allowing proliferation of effector T cells
allowing proliferation of effector T cells
Improved Output: Maintains growth of CD8⁺/CD4⁺ T cells for workflows aimed at extending proliferation of T cells.
Sustained Phenotype: Improved proliferation capacity without increased exhaustion nor more terminally differentiated T cells.

Product Specifications

Endotoxin Level

<0.20 EU per 1 μg of the protein by the LAL method.

Activity

Measured in a cell proliferation assay using NK-92 human natural killer lymphoma cells. The ED₅₀ for this effect is 0.0500-0.500 ng/mL.

Source

E. coli-derived human IL-2 protein
Ala21-Thr153 (C145S) with and without an N-terminal Met
Proprietary point mutations

Accession #

P60568.1

N-terminal Sequence
Analysis

Ala21 & Met-Ala21

Predicted Molecular Mass

15 kDa

SDS-PAGE

12-14 kDa, under reducing conditions.

Product Datasheets

BT-002DBR

Product Datasheet

COA

SDS

Carrier Free

What does CF mean?

CF stands for Carrier Free (CF). We typically add Bovine Serum Albumin (BSA) as a carrier protein to our recombinant proteins. Adding a carrier protein enhances protein stability, increases shelf-life, and allows the recombinant protein to be stored at a more dilute concentration. The carrier free version does not contain BSA.

What formulation is right for me?

In general, we advise purchasing the recombinant protein with BSA for use in cell or tissue culture, or as an ELISA standard. In contrast, the carrier free protein is recommended for applications, in which the presence of BSA could interfere.

BT-002DBR

Formulation	Lyophilized from a 0.2 μm filtered solution in Sodium Acetate with Trehalose.
Reconstitution	Reconstitute the 10 μg size at 100 μg/mL in water. Reconstitute all the other sizes at 500 μg/mL in water.
Shipping	The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage:	Use a manual defrost freezer and avoid repeated freeze-thaw cycles. 12 months from date of receipt, -20 to -70 °C as supplied. 1 month, 2 to 8 °C under sterile conditions after reconstitution. 3 months, -20 to -70 °C under sterile conditions after reconstitution.

Scientific Data

Cell Proliferation

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CD4⁺ and CD8⁺ T cells from two different donors were activated with anti-CD3-, anti-CD28-conjugated beads, and grown for 28 days in media containing either 3.6 or 14.3 ng/mL of Animal-free Recombinant Human IL-2 Protein (BT-002-AFL) as the standard IL-2 protein or engineered Recombinant Human IL-2 (CD122-Directed) Protein (BT-002DBR), refreshed every 2-3 days. The average fold expansion of the cells was determined every 2-3 days. The IL-2 (CD122-Directed) Protein supports greater than 6x more CD4+/CD8+ T cell growth than the standard IL-2 protein.

Phenotyping

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CD4⁺ and CD8⁺ T cells from three different donors were activated with anti-CD3-, anti-CD28-conjugated beads, and grown in media containing 50 or 200 IU of Animal-free Recombinant Human IL-2 Protein (BT-002-AFL) as the standard IL-2 protein or Recombinant Human IL-2 (CD122-Directed) Protein (BT-002DBR), refreshed every 2-3 days. A) After 28 days, the phenotypes of the cells were compared by flow cytometry using a Brilliant Violet 711™ anti-human CD197 (CCR7) antibody and PE/Dazzle™ 594 anti-human CD45RA antibody to determine the number of CD4⁺ and CD8⁺ naive (CD45RA⁺/CCR7⁺) Tscm cells. The percentage of Tscm cells was found to be higher when the cells were expanded with the IL-2 (CD122-Directed) Protein at 14.3 ng/mL. B) The number of cells expressing the T cell exhaustion markers, PD-1, LAG-3, and TIM-3 was determined by flow cytometry. The results showed that the percentages of CD4⁺/CD8⁺ T cells that were also PD-1, LAG-3, or TIM-3 positive were equivalent or reduced with the IL-2 (CD122-Directed) Protein. These results demonstrate that the IL-2 (CD122-Directed) Protein promotes increased T cell expansion with enhanced expression of naive phenotype markers and equivalent or reduced expression of cell exhaustion markers.

Surface Plasmon Resonance (SPR)

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Recombinant Human IL 2 R beta Fc Protein (10919-2B) was captured on a Sensor Chip Protein AGL, and binding to Recombinant Human IL-2 (CD122-Directed) Protein (BT-002DBR) was measured at a concentration range between 0.097nM and 200 nM. The double-referenced sensorgram was fit to a 1:1 binding model to determine the binding kinetics and affinity, with an affinity constant of K_d=49.3 nM. Note: The IL-2 (CD122-Directed) Protein was modeled to engage only the IL-2 dimeric beta gamma receptor complex. In comparison, wild type recombinant human IL-2 (BT-002) affinity to Recombinant Human IL 2 R beta Fc Chimera (10919-2B) has a binding constant K_d=544 nm.

Bioactivity

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Recombinant Human IL-2 (CD122-Directed) Protein (Catalog # BT-002DBR) stimulates proliferation of NK-92 human natural killer lymphoma cells. The ED₅₀ for this effect is 0.0500-0.500 ng/mL.

SDS-PAGE

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2 μg/lane of Recombinant Human IL-2 (CD122-Directed) Protein (Catalog # BT-002DBR) was resolved with SDS-PAGE under reducing (R) and non-reducing (NR) conditions and visualized by Coomassie® Blue staining, showing bands at 12‑14 kDa, under reducing conditions.

Reconstitution Calculator

Background: IL-2

CD122 directed IL-2 has a 7-fold increased binding affinity to the human IL-2 beta receptor while exhibiting significantly reduced affinity for the IL-2 alpha receptor. According to research by Levin (2012), this configuration results in more effective expansion of effector cells and less Treg (1). Our findings show that CD122 directed IL-2 improves immune cell expansion for workflows requiring significant amounts of IL-2. One concern with improving T cell expansion is the possibility of increasing the number of terminally differentiated and exhausted T cells with reduced anti-tumor activity. However, our studies have shown that CD122 directed IL-2 significantly improves T cell expansion without changing T cell phenotype or the expression of exhaustion markers.

References

Levin, A., Bates, D., Ring, A. et al. (2012) Nature. 484:529.

Long Name

Interleukin 2

Entrez Gene IDs

3558 (Human); 16183 (Mouse); 116562 (Rat); 396868 (Porcine); 280822 (Bovine); 403989 (Canine); 100034204 (Equine); 751114 (Feline); 100302458 (Rabbit)

Alternate Names

Aldesleukin; IL2; IL-2; IL-2lymphokine; interleukin 2; interleukin-2; involved in regulation of T-cell clonal expansion; Proleukin; T cell growth factor; T-cell growth factor; TCGF