Anti pd l1 clone 22c3

Manufactured by Merck Group

Sourced in United States

The Anti-PD-L1 clone 22C3 is a laboratory reagent used for the detection and analysis of programmed death-ligand 1 (PD-L1) expression in biological samples. It is a monoclonal antibody that specifically binds to the PD-L1 protein. This reagent can be utilized in various immunoassay techniques, such as flow cytometry and immunohistochemistry, to assess the levels of PD-L1 expression in cells or tissue samples.

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Lab products found in correlation

Flex polymer system, by Agilent Technologies (2 mentions) Optiview amplification kit, by Roche (1 mentions) Haematoxylin and bluing reagent, by Roche (1 mentions) Cc1 antigen retrieval solution, by Roche (1 mentions) Optiview dab ihc kit, by Roche (1 mentions) Benchmark ultra, by Roche (1 mentions) 3 3 diaminobenzidine chromogen, by Agilent Technologies (1 mentions) Envision flex target retrieval solution high ph, by Agilent Technologies (1 mentions)

5 protocols using anti pd l1 clone 22c3

Detecting Merkel Cell Carcinoma Biomarkers

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Patients were determined to have MCPyV-positive tumors if they produced small T-antigen-specific serum antibodies21 (link) or manifested large T-antigen expression in tumor biopsies via immunohistochemistry.22 (link) PD-L1 staining (anti-PD-L1 clone 22C3, Merck & Co, Kenilworth, New Jersey, USA) was performed at QualTek Molecular Laboratories on pretreatment tumor specimens as previously described.17 (link) Specimens were considered PD-L1 positive if ≥1% of tumor cells expressed PD-L1 at the cell surface.16 20 (link)
Neutrophil-to-lymphocyte ratio (NLR) was calculated using absolute neutrophil counts and absolute lymphocyte counts (ALC), determined from automated complete blood counts in peripheral blood specimens obtained at study visits. NLR was calculated at baseline (before initial pembrolizumab infusion) and after each of the first four treatment cycles.

Nghiem P., Bhatia S., Lipson E.J., Sharfman W.H., Kudchadkar R.R., Brohl A.S., Friedlander P.A., Daud A., Kluger H.M., Reddy S.A., Boulmay B.C., Riker A., Burgess M.A., Hanks B.A., Olencki T., Kendra K., Church C., Akaike T., Ramchurren N., Shinohara M.M., Salim B., Taube J.M., Jensen E., Kalabis M., Fling S.P., Homet Moreno B., Sharon E., Cheever M.A, & Topalian S.L. (2021). Three-year survival, correlates and salvage therapies in patients receiving first-line pembrolizumab for advanced Merkel cell carcinoma. Journal for Immunotherapy of Cancer, 9(4), e002478.

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Immunohistochemical Evaluation of PD-L1 Expression

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Four µm sections were cut from FFPE tissue blocks. After deparaffinization, antigen retrieval was performed by CC1 antigen retrieval solution (pH 8.0, Ventana Medical systems, Tuczon, AZ, USA) on the Ventana BenchMark Ultra automated slide stainer. Slides were incubated with the primary PD-L1 antibody (anti-PD-L1, clone 22C3, Merck) at a dilution of 1/100 for 32 min, followed by visualization with the OptiView DAB IHC Kit and OptiView Amplification Kit (Ventana Medical systems). The specimens were then counterstained with haematoxylin and bluing reagent (Ventana Medical systems) and coverslipped.
PD-L1 expression was scored in immune cells in the paracortex and in the sinuses of the lymph node, excluding the germinal centers. Similar to IDO1 scoring, the intensity of PD-L1 staining in the paracortex was evaluated according to a four-tiered grading system (Supplementary Figure 3A): no expression (0), weak expression (1+), moderate expression (2+) or strong expression (3+). PD-L1 expression in the paracortex was dichotomized into an PD-L1-low group (0 and 1+) and an PD-L1-high group (2+ and 3+). In the sinuses, PD-L1 staining was scored as ‘low’ or ‘high’ (Supplementary Figure 3B).

Meireson A., Ferdinande L., Haspeslagh M., Hennart B., Allorge D., Ost P., Sundahl N., Spaas M., Demeyer A, & Brochez L. (2021). Clinical Relevance of Serum Kyn/Trp Ratio and Basal and IFNγ-Upregulated IDO1 Expression in Peripheral Monocytes in Early Stage Melanoma. Frontiers in Immunology, 12, 736498.

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Metastatic Tumor TILs and PD-L1 Assessment

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Available pre-treatment metastatic tumor samples were assessed for TILs density, according to the suggested international guidelines [19 (link)]. Cut-offs explored for TILs evaluation were ≥ 5%, ≥ 10%, and ≥ 20%. PD-L1 expression measured using IHC was assessed in 29 pre-treatment metastatic tumor samples using the mAb anti-PD-L1 clone 22C3 (Merck, Kenilworth, NJ). We obtained combined positive scores (CPS) for the assessment of PD-L1 positivity and considered CPS values ≥ 1 as positive (PD-L1 +). We also explored additional CPS cut-offs (≥ 5, ≥ 10, and ≥ 20).
The logistic and Cox regression models were used to evaluate the association of TILs density and PD-L1 expression with treatment efficacy in terms of ORR, CBR (with SD ≥ 24 weeks) and PFS according to the RECIST 1.1.

de la Cruz-Merino L., Gion M., Cruz J., Alonso-Romero J., Quiroga V., Moreno F., Andrés R., Santisteban M., Ramos M., Holgado E., Cortés J., López-Miranda E., Cortés A., Henao F., Palazón-Carrión N., Rodriguez L.M., Ceballos I., Soto A., Puertes A., Casas M., Benito S., Chiesa M., Bezares S., Caballero R., Jiménez-Cortegana C., Sánchez-Margalet V, & Rojo F. (2022). Pembrolizumab in combination with gemcitabine for patients with HER2-negative advanced breast cancer: GEICAM/2015–04 (PANGEA-Breast) study. BMC Cancer, 22, 1258.

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IHC Scoring of PD-L1 and PD-1 Biomarkers

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Formalin‐fixed, paraffin‐embedded (FFPE) tissue blocks were deparaffinized and rehydrated with serial passage through changes of xylene and graded ethanols for PD‐L1 and PD‐1 IHC. All slides were subjected to heat‐induced epitope retrieval in Envision FLEX Target Retrieval Solution, High pH (Dako Corporation, Carpinteria, Calif). Endogenous peroxidase in tissues was blocked by incubation of slides in 3% hydrogen peroxide solution before incubation with the primary antibody (anti‐PD‐L1 clone 22C3 [Merck Research Laboratories, Palo Alto, Calif] or anti‐PD‐1 clone NAT105 [Cell Marque, Rocklin, Calif]) for 60 minutes. Antigen‐antibody binding was visualized with the FLEX + polymer system (Dako) and application of 3,3' diaminobenzidine chromogen (Dako). Stained slides were counterstained with hematoxylin and coverslipped for review and scoring. Scoring was conducted by a pathologist (J.H.Y.) who was blinded to all patient clinical information using a semiquantitative scale of 0 to 5, in which positive cell frequency within the tumor tissue was grouped into the following categories: 0 indicates negative, 1 indicates rare, 2 indicates low, 3 indicates moderate, 4 indicates high, and 5 indicates very high. These patterns are illustrated for both PD‐1 and PD‐L1 in Supporting Information Figure 1.

Pollack S.M., He Q., Yearley J.H., Emerson R., Vignali M., Zhang Y., Redman M.W., Baker K.K., Cooper S., Donahue B., Loggers E.T., Cranmer L.D., Spraker M.B., Seo Y.D., Pillarisetty V.G., Ricciotti R.W., Hoch B.L., McClanahan T.K., Murphy E., Blumenschein W.M., Townson S.M., Benzeno S., Riddell S.R, & Jones R.L. (2017). T‐cell infiltration and clonality correlate with programmed cell death protein 1 and programmed death‐ligand 1 expression in patients with soft tissue sarcomas. Cancer, 123(17), 3291-3304.

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PD-L1 and PD-1 Immunohistochemistry in FFPE Samples

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FFPE blocks were deparaffinized and rehydrated with serial passage
through changes of xylene and graded ethanols for PD-L1 and PD-1
immunohistochemistry (IHC). All slides were subjected to heat induced epitope
retrieval in Envision FLEX Target Retrieval Solution, High pH (cat K8012, Dako,
Carpineteria CA). Endogenous peroxidase in tissues was blocked by incubation of
slides in 3% hydrogen peroxide solution prior to incubation with primary
antibody (anti-PD-L1 clone 22C3, Merck Research Laboratories, Palo Alto CA or
anti-PD-1 clone NAT105, Cell Marque, Rocklin CA) for 60 minutes.
Antigen-antibody binding was visualized with the FLEX+ polymer system
(cat K8012, Dako, Carpineteria CA) and application of 3,3′
diaminobenzidine (DAB) chromogen (K4368, Dako). Stained slides were
counterstained with hematoxylin and cover-slipped for review and scoring.
Scoring was conducted by a pathologist (JHY) blinded to all patient clinical
information using a semi-quantitative 0–5 scale, in which positive cell
frequency within the tumor tissue was grouped into the following categories: 0
= negative, 1 = rare, 2 = low, 3 = moderate, 4
= high, and 5 = very high. These patterns are illustrated for
both PD-1 and PD-L1 in Supplemental Figure 1.

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