Sp142

Manufactured by Roche

Sourced in United States, Switzerland

The SP142 is a laboratory instrument designed for the analysis and quantification of specific analytes or molecules in a given sample. It utilizes advanced spectrophotometric techniques to provide accurate and reliable results. The core function of the SP142 is to perform precise measurements and data analysis, assisting researchers and scientists in their laboratory workflow.

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42 protocols using sp142

Evaluating PD-L1 Expression in Cancer-Associated Fibroblasts

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Immunohistochemical stainings were performed using autostainers (SP142 and E1L3N assays on Discovery ULTRA System; Roche Diagnostics, Basel, Switzerland; and 73–10 assay on Leica Bond-III; Leica Biosystems, Bannockburn, IL). Three different primary monoclonal antibodies were used to detect PD-L1: SP142 (Roche Diagnostics, Basel, Switzerland), E1L3N (Cell Signaling Technology, Danvers, MA, USA) and 73–10 (Leica Biosystems, Newcastle, UK). A minimum of two researchers independently evaluated the immunohistochemical staining results.
Spindle-shaped non-neoplastic cells in tumour stroma were morphologically recognized as CAFs, and membranous and/or cytoplasmic expression of PD-L1 in these cells was considered positive. PD-L1 expression scores of CAFs were determined based on the staining intensity and were classified into three levels (0, negative; + 1, weak; + 2, strong). As previously reported, CAF PD-L1 positivity was defined as the presence of CAFs with staining intensities of + 1 and + 2 in more than 1% of a section and positive immunoreactivity of ≥1 from the same patient [18 (link)]. In addition, PD-L1 expression in stromal TILs was defined as expression in more than 5% of TILs (TIL PD-L1-positive) [25 (link), 26 (link)].

Yoshikawa K., Ishida M., Yanai H., Tsuta K., Sekimoto M, & Sugie T. (2021). Prognostic significance of PD-L1-positive cancer-associated fibroblasts in patients with triple-negative breast cancer. BMC Cancer, 21, 239.

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Comprehensive Molecular Analysis of Tumor Samples

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Patients underwent whole-exome sequencing at the University of Michigan,
as we have previously described^{29 (link)}. T cell receptor sequencing and diversity calculation was
performed as we previously described using the immunoSEQ platform (Adaptive
Biotechnologies)^{29 (link)}.
Tumor mutational burden was estimated as: (total mutations / total covered
bases) × 10⁶, as previously described^{29 (link)}. A previously validated gene signature
for the CD8⁺ T cells function was utilized^{30 (link)}. Annotation of liver metastasis was
performed on the basis of cross-sectional imaging acquired immediately prior to
biopsies taken for sequencing. Signature scores were computed by inverse-normal
transformation of gene expression levels across the cohort followed by summation
of inverse-normal values for each sample, as previously described. PD-L1
expression was quantified on the basis of expression profiling (Cohort 5) or on
the basis of immunohistochemistry using 22C3 (Dako) or SP142 (Ventana) PD-L1
antibody staining performed in a CLIA ‘88 (Clinical Laboratory
Improvement Amendments) certified laboratory.

Yu J., Green M.D., Li S., Sun Y., Journey S.N., Choi J.E., Rizvi S.M., Qin A., Waninger J.J., Lang X., Chopra Z., Naqa I.E., Zhou J., Bian Y., Jiang L., Tezel A., Skvarce J., Achar R.K., Sitto M., Rosen B.S., Su F., Narayanan S.P., Cao X., Wei S., Szeliga W., Vatan L., Mayo C., Morgan M.A., Schonewolf C.A., Cuneo K., Kryczek I., Ma V.T., Lao C.D., Lawrence T.S., Ramnath N., Wen F., Chinnaiyan A.M., Cieslik M., Alva A, & Zou W. (2021). Liver metastasis restrains immunotherapy efficacy via macrophage-mediated T cell elimination. Nature medicine, 27(1), 152-164.

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PD-L1 Immunohistochemistry Antibody Evaluation

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Serial unstained slides (4 microns) were prepared from each block for subsequent immunohistochemistry (IHC) with the following PD-L1 antibody clones: SP142 (Ventana medical systems, Tucson, AZ, USA), SP263 (Ventana medical systems, Tucson, AZ, USA), 22C3 pharmDX (Agilent technologies, Santa Clara, CA, USA) and E1L3N (Cell Signaling Technologies, Danvers, MA, USA). Sections were stained with the SP142 and SP263 antibody clones on the Ventana Benchmark Ultra automated staining platform, E1L3N on Leica Bond-III Autostainer (Leica Biosystems, Buffalo Grove, IL, USA), and with the 22C3 clone utilized the DAKO EnVision FLEX system on a DAKO Autostainer Link 48 system (Agilent technologies, Santa Clara, CA, USA) according to the manufacturer’s instructions.

Downes M.R., Slodkowska E., Katabi N., Jungbluth A.A, & Xu B. (2019). Interobserver and Intraobserver agreement of PD-L1 scoring in Head and Neck Squamous Cell Carcinoma (HSCC), Urothelial Carcinoma (UC), and Breast Carcinoma (BC). Histopathology, 76(2), 191-200.

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Comprehensive PD-L1 Tumor Cell Evaluation

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All cases included in this study must also have been tested with a commercially available PD-L1 IHC assay (ie, DAKO 22C3, DAKO 28–8, VENTANA SP142, or VENTANA SP263) in addition to the CGP. The tumor cell staining (TS) score where # PD-L1 positive tumor cells / (total # of PD-L1 positive+PD-L1 negative tumor cells) was determined.

Huang R.S., Carbone D.P., Li G., Schrock A., Graf R.P., Zhang L., Murugesan K., Ross J.S., Tolba K., Sands J., Oxnard G.R, & Spigel D. (2023). Durable responders in advanced NSCLC with elevated TMB and treated with 1L immune checkpoint inhibitor: a real-world outcomes analysis. Journal for Immunotherapy of Cancer, 11(1), e005801.

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PD-L1 expression in FFPE samples

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The specimens were formalin-fixed, paraffin embedded (FFPE), sliced into 4 μm sections and stained for PD-L1 with a rabbit monoclonal antibody SP142 (Ventana/Roche, USA) on an automated platform (Benchmark, Ventana/Roche, USA). Three independent investigators (I.K., U.J. and L.C.) examined whole slices without prior knowledge of the clinicopathological features of the patients. The presence of IC (yes/no) was evaluated for each individual specimen. Percentage of PD-L1 positive immunohistochemical reaction was evaluated in TC and IC, ranging from 0–100%, regardless of staining intensity. The staining of TC on the cell membrane was regarded as positive, whereas IC showed PD-L1 positive reaction in the cytoplasm. Human placenta was also immunostained as a control tissue for PD-L1 expression. The results were then statistically analyzed for two preplanned cut-off values, namely 5% or higher and 10% or higher PD-L1 expression in either TC or IC.

Janzic U., Kern I., Janzic A., Cavka L, & Cufer T. (2017). PD-L1 Expression in Squamous-cell Carcinoma and Adenocarcinoma of the Lung. Radiology and Oncology, 51(3), 357-362.

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PD-L1 and EGFR Expression in Cancer

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Hematoxylin and eosin-stained slides and immunostained slides upon primary diagnosis (performed at the Department of Pathology in our hospital) were available for all patients. For immunostaining for SP142 (Ventana Medical Systems Inc.), the specimens were formalin-fixed, paraffin-embedded, sectioned, and stained in accordance with standard clinical operating procedures. The proportion of PD-L1-positive tumor cells was determined by calculating the percentage of stained tumor cells. Positivity was defined as a proportion of PD-L1-positive tumor cells >1%. Proportions of PD-L1-positive tumor cells ≥50% and 1–49% were defined as indicative of high and low expression, respectively.
We retrospectively reviewed the mutational status of the epidermal growth factor receptor (EGFR, exons 18–22) gene in all patients. Tumor samples were obtained through surgical resection or biopsy. EGFR mutations were analyzed using the amplification-refractory mutation system. Cycle sequencing of the purified PCR products was performed using the ADx Mutation Detection Kits (Amory, Xiamen, China) in accordance with the manufacturer's instructions.

Zhao L., Liu J., Shi J, & Wang H. (2020). Relationship between SP142 PD-L1 Expression and 18F-FDG Uptake in Non-Small-Cell Lung Cancer. Contrast Media & Molecular Imaging, 2020, 2010924.

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PD-L1 Expression in Oncology Trials

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The primary endpoint of this substudy was OS. Secondary endpoints were investigator-assessed progression-free survival (PFS) RECIST version 1.1 and objective response rate (ORR, complete/partial response) assessed by RECIST version 1.1 and modified RECIST. Immunohistochemical staining of PD-L1 expression was carried out on formalin-fixed, paraffin-embedded tissues using VENTANA SP142 rabbit monoclonal antibody (Ventana BenchMark ULTRA reader).¹⁵ (link)

Bamias A., Merseburger A.S., Loriot Y., James N., Choy E., Castellano D., Lopez-Rios F., Calabrò F., Kramer M., de Velasco G., Zakopoulou R., Tzannis K, & Sternberg C.N. (2021). SAUL, a single-arm study of atezolizumab for chemotherapy-pretreated locally advanced or metastatic carcinoma of the urinary tract: outcomes by key baseline factors, PD-L1 expression and prior platinum therapy. ESMO Open, 6(3), 100152.

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PD-L1 Expression in Refractory Lymphomas

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Formalin-fixed paraffin-embedded tissues from 78 patients with refractory and/or resistant lymphomas of B- and T-cell lineages were investigated for the expression of PD-L1 (Clones: SP142 and SP263, Ventana Medical Systems, Tucson, AZ) using automated immunohistochemical methods at a CLIA/CAP/ISO15189—certified lab (Caris Life Sciences, Phoenix, AZ). Cases were considered positive when ≥5% of the neoplastic cells exhibited membranous positivity with 2+/3+ intensity [24 (link)]. PD-L1 expression was also evaluated in adjacent population of reactive/inflammatory cells.
Two pathologists (Z.G. and S.V.) evaluated the IHC results independently; in a case of discrepant data, the cases were reviewed together and consensus was obtained.

Vranic S., Ghosh N., Kimbrough J., Bilalovic N., Bender R., Arguello D., Veloso Y., Dizdarevic A, & Gatalica Z. (2016). PD-L1 Status in Refractory Lymphomas. PLoS ONE, 11(11), e0166266.

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Tumor Biomarkers in Cancer Immunotherapy

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PD-L1 expression was assessed by immunohistochemistry (IHC) staining with an
anti-human PD-L1 monoclonal antibody (SP142, VENTANA, USA).^{12 (link)} PD-L1-positive status was defined as membrane staining of any intensity
in ⩾1% of tumor cells or tumor-infiltrating immune cells. The EBV DNA copy
number was assessed by unique reads detected in pre-treatment archived tumor
tissues. TMB was detected by whole exome sequencing with IDT xGen Exome Research
Panel version 1.0 on tumor tissues and matched peripheral blood mononuclear cell
samples. TMB was determined by analyzing somatic mutations, including coding
base substitution and INDELs per Mb. The top 20% of the TMB (12 mutations/Mb) in
this research was selected as the cut-off value and defined as
TMB^high. Patients with TMB < 12 mutations/Mb were defined as
TMB^low.

Wei X.L., Xu J.Y., Wang D.S., Chen D.L., Ren C., Li J.N., Wang F., Wang F.H, & Xu R.H. (2021). Baseline lesion number as an efficacy predictive and independent prognostic factor and its joint utility with TMB for PD-1 inhibitor treatment in advanced gastric cancer. Therapeutic Advances in Medical Oncology, 13, 1758835921988996.

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Comprehensive PD-L1 Immunohistochemistry

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Serial unstained slides (4 microns) were prepared from each block for subsequent immunohistochemistry (IHC) with the following PD-L1 antibody clones: SP142 and SP263 (Ventana Medical Systems, AZ, USA), 22C3 (Agilent Technologies, CA, USA) and E1L3N (Cell Signaling Technologies, MA, USA). Staining for anti PD-L1 rabbit mAb clones SP142, SP263 and E1L3N was performed on the Ventana Benchmark Ultra automated staining platform according to the manufacturer’s protocol (with the Optiview DAB IHC detection kit for all the clones, followed by the Optiview amplification kit for SP142 only; for mouse mAb clone 22C3, the DAKO EnVision FLEX system on a DAKO Autostainer Link 48 system was used according to the manufacturer’s instructions (Figure 1).

Hodgson A., Slodkowska E., Jungbluth A., Liu S.K., Vesprini D., Enepekides D., Higgins K., Katabi N., Xu B, & Downes M.R. (2018). PD-L1 immunohistochemistry assay concordance in urothelial carcinoma of the bladder and hypopharyngeal squamous cell carcinoma. The American journal of surgical pathology, 42(8), 1059-1066.

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