rHu IL-2 outfitted for closed-system manufacturing

Akron’s Recombinant Human Interleukin-2 (rHu IL-2) Closed System Solutions™ (CSS) is manufactured following all relevant cGMP guidelines for ancillary materials. The rHu IL-2 active substance is supported by a Type II Master File (MF) on file with the FDA and an MF Type I on file with Health Canada, which can be referenced during your drug or biologic application process.

Our rHu IL-2 amino acid sequence is identical to Proleukin® (aldesleukin), and its functional similarity in T Cell expansion has been evaluated and confirmed. Akron’s rHu IL-2 is a single chain, 15.3 kDa, non-glycosylated lymphokine analog expressed in E. coli, containing 133 amino acids. The downstream purification process uses a multi-step orthogonal approach, without the use of affinity tags, to minimize exogenous impurities and ensure the delivery of highly purified and active substance for further manufacturing applications.

The liquid rHu IL-2 CSS product is packaged in a sterile fluoropolymer bag with two different weldable tubing connection options, allowing for easy incorporation into modern closed-system cell culture bioprocessing protocols. rHu IL-2 CSS increases safety and ease of use by eliminating the reconstitution step during manufacture and allows for the introduction of cytokine material into culture media in a fully contained manner to maximize sterility. Sterile filtration and aseptic filling are followed by Endotoxin and Sterility testing performed per USP/EP on the final product. See product Packaging features below.

Active Substance

• Amino acid sequence identical to Proleukin® / aldesleukin
• Type II eCTD MF (#026152) on file with FDA and MF Type I (#e250089) on file with Health Canada
• Carrier protein-free formulation
• All raw materials are compliant, controlled, and traceable under Akron’s Quality Management System (QMS)

Manufacturing

• Multi-step downstream purification strategy excluding affinity tags
• Commercial-scale production capacity
• Sterile filtration and aseptic filling

Packaging

• Sterile bag chamber – 8-mil fluoropolymer film providing inert bio-reactivity, structural integrity, and minimal gas transmission
• Two-part tube weldable tube – top 6” portion made from weldable TPE AdvantaFlex® (1/8” ID x 1/4” OD), and the bottom 6” portion made from standard weldable PVC (3/32” ID x 5/32” OD)
• Primary packaging is plasticizer free
• Primary packaging materials extensively validated, controlled, and qualified to ensure a consistent experience
• Every bag packed in individual protective cassette and shipped in validated CSafe Parcel insulated shipper

Quality

• Relevant cGMP guidelines used in manufacture, testing, and release
– USP <1043>, Ancillary Materials for Cell, Gene, and Tissue-Engineered Products
– EP 5.2.12, Raw Materials of Biological Origin for the Production of Cell-based and Gene Therapy Medicinal Products
– ISO 13485:2016, Medical Devices – Quality Management Systems – Requirements for Regulatory Purposes
– ISO/TS 20399-1-3:2018, Biotechnology – Ancillary Materials Present During the Production of Cellular Therapeutic Products

Stability

• Under long-term stability program
• Store at 2-8 °C
• Transport with cold packs
• Do not freeze

Methods of Use

There are two different options available for connecting to and extracting the liquid solution from Akron’s rHu IL-2 CSS product (for full instructions, see “Methods of Use” document):
1) Weldable connection via top TPE AdvantaFlex® section (1/8” ID x 1/4” OD)
2) Weldable connection via bottom PVC section (3/32” ID x 5/32” OD)

For Use Statement

For research use or further manufacturing use in ex vivo cell therapy applications only. This product is not intended for direct in vivo use or for direct clinical use as a drug, therapeutic, biologic, or medical device.

Akron rHu IL-2 vs Proleukin® – pSTAT5 Expression

Figure 1: To examine IL-2 signaling, freshly isolated PBMC from healthy donors were stained with surface antibodies targeting 21 different protein markers prior to in vitro stimulation with IL-2. Single mass cell cytometry was used to compare the effect of Akron’s rHu IL-2 (top) against Proleukin® (bottom) on the expression of pSTAT5, pSTAT3, & pSTAT1 in T cell subsets. (pSTAT5 example shown above).

When results were summarized for six healthy donors, T cell stimulation in vitro by Proleukin® and Akron rHu IL-2 were indistinguishable.

This study was done in collaboration with the Dana-Farber Cancer Institute; study and poster available upon request.