Available assays

 

FDA-approved assays

 

  • Diagnostic assays can detect PD-L1 expression levels to help determine a patient's eligibility for anti–PD-1/PD-L1 therapy1-5
  • PD-L1 expression is measured across a continuum6 ; scoring cutoffs vary by assay7-10

 

Ventana PD-L1

(SP263)7

Ventana PD-L1

(SP142)8

Dako PD-L1 IHC 22C3

pharmDx9

Dako PD-L1 IHC 28-8

pharmDx10

Under clinical investigation

for NSCLC* 

Approved for use in NSCLC

Approved for use in NSCLC

Approved for use in NSCLC

Ventana PD-L1 (SP263)

Under clinical investigation for NSCLC* 

Ventana PD-L1 (SP142)8

Approved for use in NSCLC

Dako PD-L1 IHC 22C3 pharmDx9

Approved for use in NSCLC

Dako PD-L1 IHC 28-8 pharmDx10

Approved for use in NSCLC

*FDA approved for detecting PD-L1 expression in urothelial carcinoma.11 

 

 

FDA-approved assays
FDA-approved assays

 

Image reproduced with permission from Scheel AH et al 2018.12

 

The Blueprint Project found that the Ventana PD-L1 (SP263), Dako PD-L1 IHC 22C3 pharmDx, and Dako PD-L1 IHC 28-8 pharmDx assays have similar TC staining. The Ventana PD-L1 (SP142) assay showed less sensitivity.13 

 

See the product label for more information on proper use of FDA-approved companion diagnostics.

 

Laboratory-developed tests

 

LDTs were developed and their performance characteristics determined by specific institutions in a manner consistent with the Clinical Laboratory Improvement Amendments (CLIA) of 1988 requirements.14,15,†

Sample requirements vary for each LDT. Always verify that your samples adhere to the individual test specifications.14,15

These tests are performed in laboratories that adhere to performance specifications established by the Clinical Laboratory Improvement Amendments (CLIA) of 1988 under the regulation of the Centers for Medicare & Medicaid Services. They are not FDA approved.

 

 

Scoring Assays

 

Step 1: Determine the tumor cell area

 

Only certain types of cells should be used to assess PD-L1 positivity.7-10

 

Scoring Assays: Determine the tumor cell area
Scoring Assays: Determine the tumor cell area

 

Magnification 4x. Image reproduced with permission from Roche®.17

 

 

TCs (outlined in blue) should be assessed7-10

 

ICs (outlined in black) are sometimes assessed, depending on the assay7-10

 

Necrotic cells (outlined in green) should be excluded9,10,16,17

•  Normal cells should be excluded9,10

 

Step 2: Estimate the percentage of relevant cells that express PD-L1

 

Requirements and cutoffs for scoring vary by assay.7-10

 

Scoring cutoffs for PD-L1 assays in NSCLC
Scoring cutoffs for PD-L1 assays in NSCLC

 

Ventana PD-L1 (SP263) Assay7,17

 

Ventana PD-L1 (SP263) Assay – PD-L1 negative
Ventana PD-L1 (SP263) Assay – PD-L1 negative

 

Magnification 20x. Image reproduced with permission from Roche ®.17

 

PD-L1 negative

<25% of TCs have membrane PD-L1 staining 

Ventana PD-L1 (SP263) Assay – PD-L1 positive
Ventana PD-L1 (SP263) Assay – PD-L1 positive

 

 

 

PD-L1 positive

≥25% of TCs have membrane PD-L1 staining at any intensity

 

Ventana PD-L1 (SP142) Assay8,16

 

Ventana PD-L1 (SP142) Assay – PD-L1 negative
Ventana PD-L1 (SP142) Assay – PD-L1 negative

 

Magnification 20x. Image reproduced with permission from Roche ®.16

 

PD-L1 negative

<50% of TCs and <10% of ICs have PD-L1 staining 

Ventana PD-L1 (SP142) Assay – PD-L1 positive with more than 50% of TCs having membrane PD-L1 tumor staining
Ventana PD-L1 (SP142) Assay – PD-L1 positive with more than 50% of TCs having membrane PD-L1 tumor staining

 

 

 

PD-L1 positive

≥50% of TCs have membrane PD-L1 staining at any intensity

Ventana PD-L1 (SP142) Assay – PD-L1 positive with more than 10% of tumor area containing ICs with PD-L1 staining
Ventana PD-L1 (SP142) Assay – PD-L1 positive with more than 10% of tumor area containing ICs with PD-L1 staining

 

 

 

PD-L1 positive

≥10% of tumor area contains ICs with PD-L1 staining at any intensity

 

Dako PD-L1 IHC 22C3 pharmDx9

 

Dako PD-L1 IHC 22C3 pharmDx – PD-L1 negative
Dako PD-L1 IHC 22C3 pharmDx – PD-L1 negative

 

Magnification 20x. Image reproduced with permission from Agilent ®.9

 

PD-L1 negative

<1% of TCs have membrane PD-L1 staining 

Dako PD-L1 IHC 22C3 pharmDx – Low PD-L1 expression
Dako PD-L1 IHC 22C3 pharmDx – Low PD-L1 expression

 

 

 

Low PD-L1 expression

1% to 49% of TCs have membrane PD-L1 staining at any intensity

Dako PD-L1 IHC 22C3 pharmDx – High PD-L1 expression
Dako PD-L1 IHC 22C3 pharmDx – High PD-L1 expression

 

 

 

High PD-L1 expression

≥50% of TCs have membrane PD-L1 staining at any intensity

 

Dako PD-L1 IHC 28-8 pharmDx10

 

Dako PD-L1 IHC 28-8 pharmDx – PD-L1 negative
Dako PD-L1 IHC 28-8 pharmDx – PD-L1 negative

 

Image reproduced with permission from Agilent ®.10

 

PD-L1 negative

<1% of TCs have membrane PD-L1 staining 

Dako PD-L1 IHC 28-8 pharmDx – PD-L1 positive with more than 1% of TCs having membrane PD-L1 staining
Dako PD-L1 IHC 28-8 pharmDx – PD-L1 positive with more than 1% of TCs having membrane PD-L1 staining

 

 

 

PD-L1 positive

≥1% of TCs have membrane PD-L1 staining

Dako PD-L1 IHC 28-8 pharmDx – PD-L1 positive with more than 5% of TCs having membrane PD-L1 staining
Dako PD-L1 IHC 28-8 pharmDx – PD-L1 positive with more than 5% of TCs having membrane PD-L1 staining

 

 

 

PD-L1 positive

≥5% of TCs have membrane PD-L1 staining at any intensity

Dako PD-L1 IHC 28-8 pharmDx – PD-L1 positive with more than 10% of TCs having membrane PD-L1 staining
Dako PD-L1 IHC 28-8 pharmDx – PD-L1 positive with more than 10% of TCs having membrane PD-L1 staining

 

 

 

PD-L1 positive 

≥10% of TCs have membrane PD-L1 staining at any intensity

PD-L1 negative

 

<25% of TCs have membrane PD-L1 staining

 

PD-L1 positive

 

≥25% of TCs have membrane PD-L1 staining at any intensity

 

Magnification 20x. Image reproduced with permission from Roche®.17

PD-L1 negative

 

<50% of TCs and <10% of ICs have PD-L1 staining

 

PD-L1 positive

 

≥50% of TCs have membrane PD-L1 staining at any intensity

 

PD-L1 positive

 

≥10% of tumor area contains ICs with PD-L1 staining at any intensity

 

Magnification 20x. Image reproduced with permission from Roche®.16

PD-L1 negative

 

<1% of TCs have membrane PD-L1 staining

 

Low PD-L1 expression

 

1% to 49% of TCs have membrane PD-L1 staining at any intensity

 

High PD-L1 expression

 

≥50% of TCs have membrane PD-L1 staining at any intensity

 

Magnification 20x. Image reproduced with permission from Agilent®.9

PD-L1 negative

 

<1% of TCs have membrane PD-L1 staining

 

PD-L1 positive

 

≥1% of TCs have membrane PD-L1 staining 

 

PD-L1 positive

 

≥5% of TCs have membrane PD-L1 staining at any intensity

 

PD-L1 positive

 

≥10% of TCs have membrane PD-L1 staining at any intensity

 

Image reproduced with permission from Agilent®.10

 

 

PD-L1 expression is not binary; it is expressed across a continuum.6

Appropriately scoring PD-L1 expression is critical for informing treatment decisions.1-5

Appropriately scoring PD-L1 expression is critical for informing treatment decisions.1-5

lightbolb
Test your knowledge

 

 

How would you score PD-L1 expression in the following examples? 

 

Stained using Ventana PD-L1 (SP263). TCs are outlined in blue, ICs are outlined in black.17

Stained using Ventana PD-L1 (SP263). TCs are outlined in blue, ICs are outlined in black.17

PD-L1 Knowledge Test Example 1
PD-L1 Knowledge Test Example 1

Image reproduced with permission from Roche®.17

Stained using Ventana PD-L1 (SP263). TCs are outlined in blue, ICs are outlined in black.17

Stained using Ventana PD-L1 (SP263). TCs are outlined in blue, ICs are outlined in black.17

 

Stained using Ventana PD-L1 (SP263). TCs are outlined in blue, ICs are outlined in black.17

PD-L1 Knowledge Test Example 2
PD-L1 Knowledge Test Example 2

Image reproduced with permission from Roche®.17

 

 

 

Stained using Ventana PD-L1 (SP263). TCs are outlined in blue, ICs are outlined in black, necrotic cells are outlined in green.17

 

Stained using Ventana PD-L1 (SP263). TCs are outlined in blue, ICs are outlined in black, necrotic cells are outlined in green.17

PD-L1 Knowledge Test Example 3
PD-L1 Knowledge Test Example 3

Image reproduced with permission from Roche®.17

 

 

 

Stained using Ventana PD-L1 (SP263). TCs are outlined in blue, ICs are outlined in black, necrotic cells are outlined in green.17

 

Stained using Ventana PD-L1 (SP263). TCs are outlined in blue, ICs are outlined in black, necrotic cells are outlined in green.17

PD-L1 Knowledge Test Example 4
PD-L1 Knowledge Test Example 4

Image reproduced with permission from Roche®.17

 

 

FDA, Food and Drug Administration; IC, immune cell; IVD, in vitro diagnostic; LDT, laboratory-developed test; NSCLC, non–small cell lung cancer; PD-1, programmed cell death protein 1; PD-L1, programmed death-ligand 1; TC, tumor cell.

 

References: 1. Borghaei H et al. N Engl J Med. 2015;373(17):1627-1639. 2. Garon EB et al. N Engl J Med. 2015;372(21):2018-2028. 3. Herbst RS et al. Nature. 2014;515(7528):563-567. 4. Herbst RS et al. Lancet. 2016;387(10027):1540-1550. 5. Fehrenbacher L et al;  POPLAR Study Group. Lancet. 2016;387(10030):1837-1846. 6. Patel SP, Kurzrock R. Mol Cancer Ther. 2015;14(4):847-856. 7. Rebelatto MC et al. Diagn Pathol. 2016;11(1):95. 8. Ventana PD-L1 (SP142) Assay [package insert]. Tucson, AZ: Ventana Medical Systems, Inc.; 2017. 9. Dako PD-L1 IHC 22C3 pharmDx [interpretation manual]. Santa Clara, CA: Agilent Technologies; 2016. 10. Dako PD-L1 IHC 28-8 pharmDx [interpretation manual]. Santa Clara, CA; Agilent Technologies; 2015. 11. Ventana PD-L1 (SP263) Assay [package insert]. Tucson, AZ: Ventana Medical Systems, Inc.; 2017. 12. Scheel AH et al. Histopathology. 2018;72(3):449-459. 13. Tsao MS et al. J Thorac Oncol. 2018;13(9):1302-1311. 14. FDA. https://www.fda.gov/MedicalDevices/ProductsandMedicalProcedures/InVitroDiagnostics/LaboratoryDevelopedTests/default.htm. Updated October 12, 2018. Accessed November 12, 2018. 15. Fitzgibbons PL et al; College of American Pathologists Pathology and Laboratory Quality Center. Arch Pathol Lab Med. 2014;138(11):1432-1443. 16. Ventana PD-L1 (SP142) Assay [assessment guide]. Tucson, AZ: Ventana Medical Systems, Inc.; 2017. 17. Ventana PD-L1 (SP263) Assay [interpretation guide]. Tucson, AZ: Ventana Medical Systems, Inc.; 2017. 18. Lindeman NI et al. Arch Pathol Lab Med. 2013;137(6):828-860. 

 

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