Human PD-L1/B7-H1 enzyme-linked immunoassay kit

CAT: EH0048 Datasheet
Specification 96 Test
Sensitivity 8.50 pg/ml (50 μl);26.02 pg/ml (10 μl)
Standard Curve Range 40.96~10000 pg/ml
Standard Curve Gradient 7 Points
Number of Incubations 2
Sample Volume 50 μl/10 μl
Type Fully Ready-to-Use
Operation Duration 120min
pg/ml O.D. Average Corrected
0.00 0.0240 0.0272 0.0256
40.96 0.0357 0.0431 0.0394 0.0138
102.40 0.0584 0.0637 0.0611 0.0355
256.00 0.1099 0.1093 0.1096 0.0840
640.00 0.2389 0.2377 0.2383 0.2127
1600.00 0.6145 0.6434 0.6290 0.6034
4000.00 1.6350 1.7290 1.6820 1.6564
10000.00 3.6290 3.8300 3.7295 3.7039

Precision

Intra-assay Precision Inter-assay Precision
Sample Number S1 S2 S3 S1 S2 S3
22 22 22 6 6 6
Average(pg/ml) 209.2 1061.1 2965.9 236.7 1178.3 3104.7
Standard Deviation 13.1 60.9 181.0 15.6 50.0 169.9
Coefficient of Variation(%) 6.3 5.7 6.1 6.6 4.2 5.5

Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.

Inter-assay Precision (Precision between assays) Three samples of known concentration were tested six times on one plate to assess intra-assay precision.

Spike Recovery

The spike recovery was evaluated by spiking 3 levels of human PD-L1 into health human serum sample. The un-spiked serum was used as blank in this experiment.
The recovery ranged from 96% to 127% with an overall mean recovery of 117%.

Sample Values

Sample Matrix Sample Evaluated Range (pg/ml) Detectable (%) Mean of Detectable (pg/ml)
Serum 30 n.d.-124.46 63 36.72

Serum/Plasma – Thirty samples from apparently healthy volunteers were evaluated for the presence of PD-L1 in this assay. No medical histories were available for the donors.

Background: PD-L1/B7-H1

Human B7 homolog 1 (B7-H1), also called programmed death ligand 1 (PD-L1) and programmed cell death 1 ligand 1 (PDCD1L1), is a member of the growing B7 family of immune proteins that provide signals for both stimulating and inhibiting T cell activation. Other family members include B7-1, B7-2, B7-H2, PDL2 and B7-H3. B7 proteins are members of the immunoglobulin (Ig) superfamily. Their extracellular domains contain 2 Ig-like domains and all members have short cytoplasmic domains. Among the family members, there is about 20-25% amino acid identity. Human and mouse B7-H1 share approximately 70% amino acid sequence identity. B7-H1 has been identified as one of two ligands for programmed death-1 (PD-1), a member of the CD28 family of immunoreceptors. The B7-H1 gene encodes a 291 amino acid (aa) type I membrane precursor protein with a putative 18 aa signal peptide, a 220 aa extracellular domain, a 21 aa transmembrane region, and a 31 aa cytoplasmic domain. Human B7-H1 is constitutively expressed in several organs such as heart, skeletal muscle, placenta and lung, and in lower amounts in thymus, spleen, kidney and liver. B7-H1 expression is upregulated in a small fraction of activated T and B cells and a much larger fraction of activated monocytes. B7-H1 expression is also induced in dendritic cells and keratinocytes after IFN-gamma stimulation. Interaction of B7-H1 with PD-1 results in inhibition of TCR-mediated proliferation and cytokine production. The B7-H1:PD-1 pathway is involved in the negative regulation of some immune responses and may play an important role in the regulation of peripheral tolerance.

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