ELISA Kit for Programmed Cell Death Protein 1 Ligand 1 (PDL1)

CD274; PDCD1LG1; B7-H; B7H1; PD-L1; PDCD1L1; PDCD1-LG1; Programed Death Ligand 1

Specificity

This assay has high sensitivity and excellent specificity for detection of Programmed Cell Death Protein 1 Ligand 1 (PDL1).
No significant cross-reactivity or interference between Programmed Cell Death Protein 1 Ligand 1 (PDL1) and analogues was observed.

Recovery

Matrices listed below were spiked with certain level of recombinant Programmed Cell Death Protein 1 Ligand 1 (PDL1) and the recovery rates were calculated by comparing the measured value to the expected amount of Programmed Cell Death Protein 1 Ligand 1 (PDL1) in samples.

Matrix Recovery range (%) Average(%)
serum(n=5) 94-101 98
EDTA plasma(n=5) 82-98 86
heparin plasma(n=5) 80-104 89

Precision

Intra-assay Precision (Precision within an assay): 3 samples with low, middle and high level Programmed Cell Death Protein 1 Ligand 1 (PDL1) were tested 20 times on one plate, respectively.
Inter-assay Precision (Precision between assays): 3 samples with low, middle and high level Programmed Cell Death Protein 1 Ligand 1 (PDL1) were tested on 3 different plates, 8 replicates in each plate.
CV(%) = SD/meanX100
Intra-Assay: CV<10%
Inter-Assay: CV<12%

Linearity

The linearity of the kit was assayed by testing samples spiked with appropriate concentration of Programmed Cell Death Protein 1 Ligand 1 (PDL1) and their serial dilutions. The results were demonstrated by the percentage of calculated concentration to the expected.

Sample 1:2 1:4 1:8 1:16
serum(n=5) 94-102% 84-103% 99-105% 79-92%
EDTA plasma(n=5) 94-103% 97-105% 99-105% 99-105%
heparin plasma(n=5) 83-98% 85-94% 80-92% 80-94%

Stability

The stability of kit is determined by the loss rate of activity. The loss rate of this kit is less than 5% within the expiration date under appropriate storage condition.
To minimize extra influence on the performance, operation procedures and lab conditions, especially room temperature, air humidity, incubator temperature should be strictly controlled. It is also strongly suggested that the whole assay is performed by the same operator from the beginning to the end.

Reagents and materials provided

Reagents Quantity Reagents Quantity
Pre-coated, ready to use 96-well strip plate 1 Plate sealer for 96 wells 4
Standard 2 Standard Diluent 1×20mL
Detection Reagent A 1×120µL Assay Diluent A 1×12mL
Detection Reagent B 1×120µL Assay Diluent B 1×12mL
TMB Substrate 1×9mL Stop Solution 1×6mL
Wash Buffer (30 × concentrate) 1×20mL Instruction manual 1

Assay procedure summary

1. Prepare all reagents, samples and standards;
2. Add 100µL standard or sample to each well. Incubate 1 hours at 37°C;
3. Aspirate and add 100µL prepared Detection Reagent A. Incubate 1 hour at 37°C;
4. Aspirate and wash 3 times;
5. Add 100µL prepared Detection Reagent B. Incubate 30 minutes at 37°C;
6. Aspirate and wash 5 times;
7. Add 90µL Substrate Solution. Incubate 10-20 minutes at 37°C;
8. Add 50µL Stop Solution. Read at 450nm immediately.

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Magazine Citations
Chinese Journal of Cancer Level of circulating PD-L1 expression in patients with advanced gastric cancer and its clinical implications NCBI: PMC3937742
Leukemia.? High level of soluble programmed cell death ligand 1 in blood impacts overall survival in aggressive diffuse large B-Cell lymphoma: results from a French multicenter clinical trial. Pubmed:24732592
Diabetes?Metab Res Rev. PD‐L1 gene polymorphisms and low serum level of PD‐L1 protein are associated to type 1 diabetes in Chile Pubmed:24816853
Lancet Haematol Ratios of T-cell immune effectors and checkpoint molecules as prognostic biomarkers in diffuse large B-cell lymphoma: a population-based study PubMed: 26686046
Eur J Cancer.  High levels of the soluble programmed death-ligand (sPD-L1) identify hepatocellular carcinoma patients with a poor prognosis. Pubmed:27039170
Journal of Cancer Research and Clinical Oncology Serum levels of soluble programmed cell death ligand 1 as a prognostic factor on the first-line treatment of metastatic or recurrent gastric cancer article:10.1007
Journal of Hematology & Oncology PD-L1 is upregulated by EBV-driven LMP1 through NF-κB pathway and correlates with poor prognosis in natural killer/T-cell lymphoma pubmed:27737703
Lung Cancer. High plasma levels of soluble programmed cell death ligand 1 are prognostic for reduced survival in advanced lung cancer. pubmed:28212990
Oncotarget Soluble programmed death-ligand 1 (sPDL1) and neutrophil-to-lymphocyte ratio (NLR) predictssurvival in advanced biliary tract cancer patients treated with palliative chemotherapy. pubmed:27780932
Oncotarget High post-treatment serum levels of soluble programmed cell death ligand 1 predict earlyrelapse and poor prognosis in extranodal NK/T cell lymphoma patients. pubmed:27105512
Oncotarget.  Soluble PD-L1: A biomarker to predict progression of autologous transplantation in patients with multiple myeloma pubmed:27566569
Cancer Immunol Immunother. Programmed death-ligand 1 and its soluble form are highly expressed in nasal natural killer/T-cell lymphoma: a potential rationale for immunotherapy. pubmed:28349165
Leukemia Soluble programmed death-ligand 1 as a prognostic biomarker for overall survival in patients with diffuse large B-cell lymphoma: a replication study and combined analysis of 508 patients leu2016385a
Medicine Plasma levels of soluble programmed death ligand-1 may be associated with overall survival in nonsmall cell lung cancer patients receiving thoracic radiotherapy. pubmed:28207525
Allergology International  Inverse correlation of soluble programmed cell death-1 ligand-1 (sPD-L1) with eosinophil count and clinical severity in allergic rhinitis patients pubmed:27617656
Medicine Plasma levels of soluble programmed death ligand-1 may be associated with overall survival in nonsmall cell lung cancer patients receiving thoracic radiotherapy PMC5319514
American Journal of Respiratory and Critical Care Medicine Soluble Programed Death Receptor 1 Ligand (sPD-L1) in Pleural Fluid of Patients with Malignant Pleural Mesothelioma (MPM) Pdf:10.1164
Clinical Lung Cancer Soluble Programmed Cell Death Ligand 1 as a Novel Biomarker for Nivolumab Therapy for Non–Small-cell Lung Cancer Pubmed:29859759
Journal of Rheumatology Increased levels of soluble programmed death ligand 1 associate with malignancy in patients with dermatomyositis Pubmed:29419471
Translational Oncology High Serum Level of Soluble Programmed Death Ligand 1 is Associated With a Poor Prognosis in Hodgkin Lymphoma Pubmed:29698935
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