Pharmdx kit

Manufactured by Agilent Technologies

Sourced in Denmark

The PharmDx kit is a diagnostic tool designed for use in pharmaceutical laboratories. It is a comprehensive solution for the analysis and identification of pharmaceutical compounds. The kit provides a standardized and efficient approach to pharmaceutical testing, enabling accurate and reliable results.

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Lab products found in correlation

Bond 3, by Leica (1 mentions) Dako automated link 48 platform, by Agilent Technologies (1 mentions) Link 48, by Agilent Technologies (1 mentions) Pd l1 22c3 pharmdx kit, by Agilent Technologies (1 mentions) Ventana discovery xt system, by Roche (1 mentions) M7240, by Agilent Technologies (1 mentions) Discovery xt, by Roche (1 mentions)

Close spelling variants of this product

Anti-PD-L1 [22C3] pharmDx kit EGFR pharmDx kit Dako PD-L1 22C3 pharmDx kit HER2 CISH pharmDx kit PD-L1 clone 22C3 pharmDx kit Dako PD-L1 IHC 22C3 pharmDx kit TOP2A FISH pharmDx™ Kit PD-L1 22C3 pharmDx kit Dako Her2 CISH pharmDx Kit 22C3 pharmDx kit

8 protocols using pharmdx kit

NSCLC Tumor Tissue IHC Staining

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A total of 130 deidentified, archival formalin-fixed, paraffin-embedded (FFPE) tumor tissues (1 tissue sample/patient) from NSCLC patients were used in this study. All the FFPE blocks were processed, reviewed, and stored in a College of American Pathologists (CAP) accredited and Clinical Laboratory Improvement Amendments (CLIA) certified laboratory (Tempus Labs Chicago, IL, USA).
Each FFPE block was cut into 4 μm-thick serial sections for H and E and IHC stains. H and E staining was performed on the Leica Autostainer XL staining platform. The sections were stained with H and E and anti-PD-L1 (clone 22c3, pharmDx Kit Dako) using an automated staining system (BOND-III: Leica Microsystems). IHC slides were stained with anti-PD-L1 22C3 monoclonal mouse primary antibody using the Bond Polymer Refine detection system on a Leica Microsystems BOND-III with positive and negative cell line run controls.

Sha L., Osinski B.L., Ho I.Y., Tan T.L., Willis C., Weiss H., Beaubier N., Mahon B.M., Taxter T.J, & Yip S.S. (2019). Multi-Field-of-View Deep Learning Model Predicts Nonsmall Cell Lung Cancer Programmed Death-Ligand 1 Status from Whole-Slide Hematoxylin and Eosin Images. Journal of Pathology Informatics, 10, 24.

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Immunohistochemical Evaluation of EGFR, HER3, and HER4

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Immunohistochemical staining was performed on the 4-μm-thick sections of TMA blocks with EGFR (pharmDX kit, prediluted, DAKO, Glostrup, Denmark), HER3 (1:20, DAKO), and HER4 (1:8,000, Thermo Scientific, Fremont, CA) antibodies. Staining was carried out according to the manufacturer’s protocols. In brief, the slides from the TMA blocks with 4-μm thickness were deparaffinized and rehydrated. Then, these slides were placed for antigen retrieval and incubated with antibodies [13 (link),16 (link),17 (link)]. EGFR-positive expression was defined as membranous staining in > 0% of the tumor cells, according to the DAKO EGFR pharmDX assay interpretation guideline. For evaluation of HER3 and HER4 expression, a previously described 4-tiered scoring system based on the staining intensity was applied [16 (link)]. The cytoplasmic and/or membranous expression of HER3 was categorized into four grades, as follows: 0, negative; 1, weak staining; 2, moderate staining; and 3, strong staining. In addition, cytoplasmic and nuclear expression of HER4 was evaluated separately. Positivity for HER3 and HER4 expression was considered as ≥ 10% of tumor cells with moderate to strong immunostaining.

Yun S., Kwak Y., Nam S.K., Seo A.N., Oh H.K., Kim D.W., Kang S.B, & Lee H.S. (2018). Ligand-Independent Epidermal Growth Factor Receptor Overexpression Correlates with Poor Prognosis in Colorectal Cancer. Cancer Research and Treatment : Official Journal of Korean Cancer Association, 50(4), 1351-1361.

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Histopathological and IHC Analysis of Gingiva

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This procedure routinely was processed in the Histopathology Laboratory of Shorsh Hospital/Sulaymaniyah Governorate. Briefly, the gingival samples were embedded in paraffin blocks, and then, 3 sections were cut; one of them (4 μm thickness) was mounted on an ordinary slide for hematoxylin and eosin (H and E) staining for histopathological observation. On the other hand, the other two sections were mounted on positively charged slides for immunohistochemistry staining for the EGFR marker (pharmDx™ kit, Dako, Glostrup, Denmark IHC Detection System) and the P53 marker (Clone DO-7, Biogenex Life Sciences Limited, Fremont, CA, USA).

Ahmed A.G, & Raziq A.H. (2019). Evaluation of Light-Emitting Diodes’ Effects on the Expression Level of P53 and EGFR in the Gingival Tissues of Albino Rats. Medicina, 55(9), 605.

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Standardized PD-L1 IHC Quantification

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PD‐L1 immunohistochemistry (IHC) testing was performed using the PD‐L1 clone 22C3, and the pharmDx kit, and Dako Automated Link 48 platform (Dako). PD‐L1 expression measured as TPS was calculated as the percentage of positive cells in at least 100 viable tumor cells with complete or partial membrane staining assessed by four experienced pathologists (LL, MB, MAB, and GGR).
²² In instances of discrepancy, a consensus meeting involving a fifth senior pathologist was convened to reach an agreement, and the kappa coefficient was used. A PD‐L1 TPS <1% was defined as negative, and a PD‐L1 TPS ≥1% was considered positive. Additionally, PD‐L1 positive samples were stratified as low PD‐L1 expression (PD‐L1 TPS 1%–49%) and high PD‐L1 expression (PD‐L1 TPS ≥50%).

Ruiz G., Enrico D., Mahmoud Y.D., Ruiz A., Cantarella M.F., Leguina L., Barberis M., Beña A., Brest E., Starapoli S., Mendoza Bertelli A., Tsou F., Pupareli C., Coppola M.P., Scocimarro A., Sena S., Levit P., Perfetti A., Aman E., Girotti M.R., Arrieta O., Martín C, & Salanova R. (2024). Association of PD‐L1 expression with driver gene mutations and clinicopathological characteristics in non‐small cell lung cancer: A real‐world study of 10 441 patients. Thoracic Cancer, 15(11), 895-905.

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PD-L1 Expression Analysis by IHC

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For PD-L1 expression analysis, IHC using an automated stainer (Link 48, Dako) was performed on 4μm sections cut from archival FFPE tumor samples from 232 patients diagnosed with non-small cell lung cancer that were retrospectively selected from the QEII Health Sciences Centre. PD-L1 IHC using the PD-L1 22C3 pharmDx kit on the Dako platform (Product number: SK006) was performed according to manufacturer recommendations [17 (link)]. The positive and negative controls were from known PD-L1 IHC positive and negative cases confirmed by IHC testing. The pharmDx kit (Dako) is designed to perform the staining using a linker and a chromogen enhancement reagent. Pre-treatment of the slides including deparaffinization and rehydration was performed using PT Link machine. Next, the specimens were incubated with monoclonal mouse IgG antibody to PD-L1, followed by incubation with a mouse linker and with a ready-to-use Visualization Reagent consisting of Goat secondary antibodies against mouse immunoglobulin and horseradish peroxidase coupled to a dextran polymer backbone. Then, chromogen and chromogen enhancement reagents were added, resulting in a brown color at the site of the antigen-antibody interaction. All slides were cover slipped and visualized with a light microscope.

Alwithenani A., Bethune D., Castonguay M., Drucker A., Flowerdew G., Forsythe M., French D., Fris J., Greer W., Henteleff H., MacNeil M., Marignani P., Morzycki W., Plourde M., Snow S., Marcato P, & Xu Z. (2021). Profiling non-small cell lung cancer reveals that PD-L1 is associated with wild type EGFR and vascular invasion, and immunohistochemistry quantification of PD-L1 correlates weakly with RT-qPCR. PLoS ONE, 16(5), e0251080.

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Quantitative Immunohistochemical Analysis

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Expression levels of therapeutic targets such as HER2/neu, EGFR and MIB1 were determined using immunohistochemistry (IHC). The immunohistochemical stainings with anti-HER2/neu (A0785; 1:300, DAKO, Hamburg, Germany), anti-EGFR (pharmDx™-kit, DAKO) and anti-MIB1 (M7240; 1:100, DAKO) antibodies were performed on 3 μm consecutive sections using an automated slide processing system (Ventana DISCOVERY XT System, Ventana Medical Systems, Inc., Tucson, USA) in accordance with the manufacturer's instructions. IHC staining and analysis were performed as previously described [37 (link)]. MIB1 score was defined as Score 0 (0 - 20% of all nuclei were positive), Score 1 (20 – 50%), Score 2 (50 – 80 %) and Score 3 (80 - 100%).

Kunzke T., Balluff B., Feuchtinger A., Buck A., Langer R., Luber B., Lordick F., Zitzelsberger H., Aichler M, & Walch A. (2017). Native glycan fragments detected by MALDI-FT-ICR mass spectrometry imaging impact gastric cancer biology and patient outcome. Oncotarget, 8(40), 68012-68025.

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Immunohistochemical Profiling of Human Tumor Tissue

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Immunohistochemical staining and analysis of human patient material was performed on tissue microarrays (TMA) as previously described(80 (link)). All tissues analyzed in this study were obtained from patients at the Institute of Surgical Pathology, University Medical Center Freiburg, Germany. This study was approved by the Ethics Committee of the University Medical Center Freiburg (no. EK 10/12). The patient cohort (n=375) comprised adenocarcinomas (n=186) and squamous cell carcinomas (n=189). Where required, histological diagnoses were verified according to current WHO classification with immunohistochemistry(81 (link)). The maximum follow-up time was 210 months. Clinical data comprising pTMN, resection status (R), gender and patient age at diagnosis are summarized in Table S15. Immunohistochemical staining was performed on a Discovery XT automated stainer (Ventana) using the following primary antibodies: MELK (1:50, Sigma-Aldrich), DDR1 (1:50, Biozol), and EGFR (pharmDx™-kit, DAKO). Signal detection was performed using peroxidase-DAB (diaminobenzidine)-MAP kit (Roche, Ventana). The immunohistological expression of the membrane-binding EGFR, the nuclear MELK and the cytoplasmatic DDR1 were classified into four levels according to specific staining intensity: Score 0, 1+, 2+, 3+.

Klaeger S., Heinzlmeir S., Wilhelm M., Polzer H., Vick B., Koenig P.A., Reinecke M., Ruprecht B., Petzoldt S., Meng C., Zecha J., Reiter K., Qiao H., Helm D., Koch H., Schoof M., Canevari G., Casale E., Re Depaolini S., Feuchtinger A., Wu Z., Schmidt T., Rueckert L., Becker W., Huenges J., Garz A.K., Gohlke B.O., Zolg D.P., Kayser G., Vooder T., Preissner R., Hahne H., Tonisson N., Kramer K., Goetze K., Bassermann F., Schlegl J., Ehrlich H.C., Aiche S., Walch A., Greif P.A., Schneider S., Felder E.R., Ruland J., Medard G., Jeremias I., Spiekermann K, & Kuster B. (2017). The target landscape of clinical kinase drugs. Science (New York, N.Y.), 358(6367), eaan4368.

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EGFR Protein Expression Analysis

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EGFR protein expression was evaluated by immunohistochemistry using a mouse anti-human EGFR monoclonal antibody (clone 2-18C9, Pharm Dx kit, Dako North America, Inc., Via Real, Carpinteria, CA, USA), according to the manufacturer’s instructions. Sections (4 μm thick) were cut from paraffin tissue blocks, deparaffinized in xylene, and rehydrated in decreasing concentrations of ethanol. Appropriate positive and negative controls were used, and positive results were defined as samples demonstrating greater than 10% of tumor cells with membranous staining of any intensity. The percentage of positive cells and the intensity, which was defined as mild (1+), moderate (2+) or strong (3+) were recorded for each sample.

Zhang J., Wang C., Gao J., Sun J., Zeng X., Wu S, & Liang Z. (2014). Adrenal cortical neoplasms: a study of clinicopathological features related to epidermal growth factor receptor gene status. Diagnostic Pathology, 9, 19.

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