CD39/CD73: the critical targets for adenosinergic pathways

The ATP–adenosine pathway functions as a key modulator of innate and adaptive immunity within the tumour microenvironment.

CD39 and CD73, two of the Ectonucleotidases, are involved in converting ATP into extracellular adenosine. CD39 (ectonucleoside triphosphate diphosphohydrolase-1, ENTPD1) converts ATP and ADP to AMP, and CD73 (ecto-5′-nucleotidase, NT5E) converts AMP to adenosine (ADO). ADO is a key metabolite within the TME that accumulates and inhibits the function of important immune cells, including T cells and NK cells, leading to an environment conducive to tumor growth. Consequently, multiple clinical strategies are being explored to target this pathway for the treatment of cancer.

The enzymatic activities of CD39 and CD73 play strategic roles in calibrating the duration, magnitude, and chemical nature of purinergic signals delivered to immune cells through the conversion of ADP/ATP to AMP and AMP to adenosine, respectively.

ACROBiosystems supplies a series of CD39/CD73 proteins with high enzymatic activity,which can help expedite your anti-CD39 and anti-CD73 mAb drug development. The high purity and bioactivity of these proteins have been verified by HPLC / BLI / SPR and ELISA. Protocols are provided for free.

High enzymatic activity

Product	Human CD39 / ENTPD1 Protein, His Tag		Human CD73 / NT5E Protein, His Tag (HPLC-verified)
Hydrolyzed substrate	Adenosine-5’-triphosphate (ATP)		Adenosine-5’-monophosphate (AMP)
Enzymatic activity （pmol/min/μg）	ACRO product（Cat. No: CD9-H52H4）	The product of another vendor	ACRO product（Cat. No: CD3-H52H7）	The product of another vendor
	> 6,000	>5,000	>37,000	>15,000

Note: The enzymatic activity of the same product is measured under the same conditions

High purity verified by HPLC

The purity of Human CD73, His Tag (HPLC-verified) (Cat. No. CD3-H52H7) was greater than 90% as determined by SEC-HPLC.

Protocol

High bioactivity

ELISA

Immobilized Human CD39, His Tag (Cat. No. CD9-H52H4) at 1 μg/mL (100 μL/well) can bind Anti-CD39 Antibody, Human IgG1 with a linear range of 0.1-0.8 ng/mL (QC tested).

Protocol

CD73 pre-coupling magnetic beads (Cat. No. MBS-K022) can bind CD73 antibody well

Immobilized 62 μg CD73 protein to 1 mg Beads can bind the Anti-CD73 Antibody with an EC50 of 0.5659 μg/mL (QC tested).

Protocol

SPR

Anti-CD73 antibody (Human IgG1) captured on CM5 chip via Anti-human IgG Fc antibodies surface can bind Human CD73, His Tag (HPLC-verified) (Cat. No. CD3-H52H7) with an affinity constant of 0.164 nM as determined in a SPR assay (Biacore 8K) (Routinely tested).

Protocol

BLI

Loaded Anti-CD73 MAb (Human IgG1) on AHC Biosensor, can bind Human CD73, His Tag (Cat. No.CD3-H52H7) with an affinity constant of 0.179 nM as determined in BLI assay (ForteBio Octet Red96e).
 

Protocol