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Your Position: Home > Leukocyte Surface Antigen CD47

Leukocyte Surface Antigen CD47

Leukocyte Surface Antigen CD47
Background

CD47,or Integrin-associated protein (IAP), is first discovered in a study aimed  at studying integrin αvβ3. It turned out to be associated in cis with integrin αvβ3 on the cell membrane. Followup studies has established CD47 as the quintessential "marker of self" expressed in all cell types. It's a key regulator of tissue homeostasis, and is also involved in a variety of diseases ranging from atherosclerosis to cancer. CD47 protects the host cells by suppressing phagocytosis. It's well established that cancer cell-expressed CD47 interacts with the NK cell-expressed SIRP family proteins, SIRP alpha and SIRP gamma. By so doing, it protects the cancer cell from being eliminated by the innate immune response.

In addition, CD47 is also essential for TSP-1 mediated vascular NO signaling.  It regulates CD36 and VEGFR2 to inhibit angiogenesis. Long term dysregulation  of NO signaling is a hallmark for several types of cardiovascular diseases,  which makes CD47 a promissing target for atherosclerosis. Indeed, Anti-CD47 mAb  has been demonstrated to reduce the atherosclerotic burden in several in vivo  models.

CD47 proteins

Given its significant role in pathogenesis of cancer and cardiovascular diseases, there is no surprise that CD47 signaling has been heavily investigated for new targeted therapeutics. ACROBiosystems provides a unique set of CD47-relevant products, including mutilple avi tag pre-biotinylated proteins, for rapid high throughput screening. The following pairs of products can be easily adapted to ELISA, Alphalisa, and MSD platform to screen neutralizing compound.

Product List
MoleculeCat. No.HostProduct DescriptionStructure
Assay data

High homogeneity verified by HPLC

The purity of Human CD47, Fc Tag (HPLC-verified) (Cat. No. CD7-H5256) was greater than 90% as determined by SEC-HPLC.

Verification of CD47 binding to SIRP-α

> Bioactivity-ELISA: CD47 can bind to SIRP-α

Immobilized Human SIRP alpha, His Tag (HPLC-verified) (Cat. No. SIA-H5225) at 2 μg/mL (100 μL/well) can bind Biotinylated Human CD47, Fc,Avitag (Cat. No. CD7-H82F6) with a linear range of 8-250 ng/mL.

> Bioactivity-BLI: CD47 can bind to SIRP-α

Immobilized Human CD47, Fc Tag (HPLC-verified) (Cat. No. CD7-H5256) on AHC Biosensor, can bind Human SIRP alpha, His Tag (HPLC-verified) (Cat. No. SIA-H5225) with an affinity constant of 0.72 μM as determined in BLI assay (ForteBio Octet Red96e)

> Cell based assay: CD47 can bind to ACHN cell expressing SIRP-α

FACS assay shows that Human CD47, Fc Tag (HPLC-verified) (Cat. No. CD7-H5256) can bind to ACHN cell expressing SIRP-α. The concentration of CD47 used is 10 μg/mL.

Application of CD47 antibody screening

Antibody affinity testing
> Bioactivity-ELISA: CD47 can bind to Anti-CD47 MAb

Immobilized Anti-CD47 MAb, Human IgG4 at 2 μg/mL (100 μL/well) can bind Biotinylated Human CD47, His,Avitag (Cat. No. CD7-H82E9) with a linear range of 0.05-2 ng/mL

> Bioactivity-SPR/BLI: CD47 can bind to Anti-CD47 MAb

Anti-Human CD47 MAb (Human IgG4) captured on CM5 chip via Anti-Human IgG Fc antibodies surface, can bind Human CD47, His Tag (Cat. No. CD7-H5227) with an affinity constant of 1.66 nM as determined in a SPR assay (Biacore T200)

Immobilized Human CD47, Fc Tag (HPLC-verified) (Cat. No. CD7-H5256) on AHC Biosensor, can bind Human SIRP alpha, His Tag (HPLC-verified) (Cat. No. SIA-H5225) with an affinity constant of 0.72 μM as determined in BLI assay (ForteBio Octet Red96e)

Application of neutralizing antibody screening

Serial dilutions of Anti-Human CD47 Neutralizing Antibody were added into Human SIRP alpha, His Tag (HPLC-verified) (Cat. No. SIA-H5225): Biotinylated Human CD47, Fc,Avitag (Cat. No. CD7-H82F6) binding reactions. The half maximal inhibitory concentration (IC50) is 124.5 ng/mL.

FACS analysis shows that the binding of Human CD47 to ACHN expressing SIRP-a was inhibited by increasing concentration of neutralizing SIRP-a antibody with the IC50 15.02 μg/mL. The concentration of Human CD47 used is 10 μg/mL.

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