Sp263 assay

Manufactured by Roche

Sourced in United States

The SP263 assay is a laboratory equipment product designed to perform specific analytical tests. It serves as a tool for researchers and scientists in various fields. The core function of the SP263 assay is to facilitate the analysis and detection of specific target analytes or molecules, providing valuable data for research and diagnostic purposes. No further interpretation or extrapolation on the intended use of this product is provided.

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Lab products found in correlation

Clone e1l3n, by Cell Signaling Technology (1 mentions) E1l3n, by Agilent Technologies (1 mentions) Benchmark platform, by Roche (1 mentions) Benchmark ultra, by Roche (1 mentions) 22c3 pharmdx assay, by Agilent Technologies (1 mentions)

Close spelling variants of this product

SP263 (70 citations) VENTANA PD-L1 (SP263) Assay (23 citations) PD-L1 (SP263) Assay (14 citations) Ventana SP263 assay (8 citations) PD-L1 SP263 IHC assay (3 citations) VENTANA PD-L1 (SP263) immunohistochemistry assay (2 citations) VENTANA PD-L1 (SP263) IHC assay (2 citations) SP263 immunohistochemistry assay (2 citations) VENTANA SP263 immunohistochemistry assay (2 citations)

5 protocols using sp263 assay

Comprehensive PD-L1 Immunohistochemistry Assay Evaluation

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A first set of three antibodies was tested: Clone E1L3N (Cell Signaling Technology, Danvers, MA, USA), clone 22C3 (Pharm Dx kit, DAKO, Agilent Technology, Santa Clara, CA, USA) and clone SP142 (Spring Bioscience, Pleasanton, CA, USA). We completed the study with the SP263 assay (Ventana Medical System, Tucson, USA) when it became commercially available, but we could only determine the SP263 status for 83 samples. SP142 was tested both as CA on Benchmark Ultra or as LDT on Dako autostainer, E1L3N was tested as LDT on Dako autostainer. PD-L1 22C3 PharmDx assay was performed on Dako Autostainer 48 according to the manufacturer’s instructions. PD-L1 SP263 commercial assay was performed on Benchmark Ultra according to the manufacturer’s instructions. See Table 2 for further details.

Table 2

Antibodies and technical data

Antibody	Clone	Provider	Visualisation system	Dilution
PD-L1	E1L3N (LDT)	Cell Signaling Technology	Envision Flex Sytem Dako	1/500
PD-L1	22C3 (CA)	Agilent (Dako)	Envision Flex Sytem Dako	Prediluted
PD-L1	SP263 (CA)	Roche Ventana	Optiview system Ventana	Prediluted
PD-L1	SP142 (LDT)	Roche (Spring biosciences)	Envision Flex Sytem Dako	1/100
PD-L1	SP142 (CA)	Roche (Spring biosciences)	Optiview system Ventana	1/60
PD1	Nat105	Roche Ventana	Optiview system Ventana	Prediluted
CD8	SP57	Roche Ventana	Optiview system Ventana	Prediluted
PD-L2	D7U8C	Cell Signaling Technology	Envision Flex Sytem Dako	1/100

Rouquette I., Taranchon-Clermont E., Gilhodes J., Bluthgen M.V., Perallon R., Chalabreysse L., De Muret A., Hofman V., Marx A., Parrens M., Secq V., Thomas de Montpreville V., Galateau-Salle F., Brousset P., Milia J., Girard N., Besse B., Molina T.J, & Mazières J. (2019). Immune biomarkers in thymic epithelial tumors: expression patterns, prognostic value and comparison of diagnostic tests for PD-L1. Biomarker Research, 7, 28.

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Evaluating PD-L1 Expression in Cancer

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PD-L1 expression was evaluated with Ventana SP263 assay on the BenchMark platform (Ventana, Tucson, AZ, USA). The PD-L1 expression positivity means over 1% of tumor cells based on NCCN Guidelines.7 We required the samples to include at least 100 viable cancer cells, assessed by two experienced pathologists for PD-L1 immunohistochemical analysis. Thirty-nine patients’ samples were available for evaluation of PD-L1 expression.

Ma Y., Li Q., Du Y., Chen W., Zhao G., Liu X., Ye L., Li H., Wang X., Liu J., Shen Z., Ma L, & Zhou Y. (2020). Tumor Mutational Burden and PD-L1 Expression in Non-Small-Cell Lung Cancer (NSCLC) in Southwestern China. OncoTargets and therapy, 13, 5191-5198.

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Multimodal Immune Profiling in Oncology

Cited 2 times

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CD8 (C8/144B, 1:100) and PD-L1 (Ventana SP263 assay) immunohistochemistry was performed on a Ventana Benchmark Ultra autostainer according to accredited staining protocols (https://www.dakks.de/en). PD-L1 was scored according to the durvalumab-linked PD-L1 scoring algorithm (TC_area-score[%]=positive tumor cell area per total tumor cell area; IC_area-score[%]=proportion of the area occupied by PD-L1 positive tumor associated IC per total area occupied by tumor-associated IC). Whole slides stained for CD8 were digitalized (P250 slide scanner, 3DHistech), and CD8 infiltration was detected quantitatively (per mm2 (link) intratumoral (tumor cell area), in the tumor associated stroma and in the total tumor area using QuPath v0.2.0. Details were described previously.16 18 (link)
Whole blood samples of the patients collected at study inclusion were analyzed with multi-color ﬂow cytometry according to previously published and clinically applied immunophenotypin protocols.16 19 (link)

Hecht M., Eckstein M., Rutzner S., von der Grün J., Illmer T., Klautke G., Laban S., Hautmann M.G., Brunner T.B., Tamaskovics B., Hinke A., Zhou J.G., Frey B., Donaubauer A.J., Becker I., Semrau S., Hartmann A., Balermpas P., Budach W., Gaipl U.S., Iro H., Gostian A.O, & Fietkau R. (2022). Induction chemoimmunotherapy followed by CD8+ immune cell-based patient selection for chemotherapy-free radioimmunotherapy in locally advanced head and neck cancer. Journal for Immunotherapy of Cancer, 10(1), e003747.

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PD-L1 Immunohistochemistry Analysis in Tumor Tissues

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Immunohistochemistry analysis of PD‐L1 was done on pretreatment formalin‐fixed paraffin‐embedded tumor tissues using the SP263 assay (Ventana Medical Systems, Tucson, AZ, United States) or the 22C3 pharmDx assay (Dako, Carpinteria, CA, United States) according to the manufacturer's recommendations. The expression of PD‐L1 was evaluated by the percentage of tumor cells with membranous staining of PD‐L1¹⁸ and assessed by pathologists in a blinded fashion.

Kim M., Keam B., Ock C., Kim S.H., Kim Y.J., Lim S.M., Kim J., Kim T.M., Hong S., Ahn M.S., Shin S.H., Kang E.J., Kim D., Im S., Kim J., Lee J.S., Kim J, & Heo D.S. (2020). Phase II study of durvalumab and tremelimumab in pulmonary sarcomatoid carcinoma: KCSG‐LU16‐07. Thoracic Cancer, 11(12), 3482-3489.

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Tumor Microenvironment Characterization by Biopsy

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Biopsies of the injected lesions were performed before the first dose of OH2 and subsequently at each response evaluation. Three biomarkers, including programmed death-ligand 1 (PD-L1), CD8 and CD3, using immunohistochemistry (IHC) were stained to characterize the changes in the tumor microenvironment. PD-L1 expression was assessed with the Ventana SP263 assay, and was reported using both the tumor proportion score (TPS) and the combined positive score (CPS). TPS was defined as the percentage of tumor cells with membranous PD-L1 staining, whereas CPS was defined as the number of PD-L1 staining cells (tumor cells, lymphocytes and macrophages) divided by the total number of viable tumor cells, multiplied by 100. For the quantification of CD8⁺ and CD3⁺ cells, the anti-CD8 mouse monoclonal antibody clone C8/144B was used for CD8 IHC, while the CD3 IHC analysis was conducted using the anti-CD3 (2GV6) rabbit monoclonal antibody. The density of CD8⁺ and CD3⁺ stained cells in the intratumoral area plus the invasive margin, if present in the biopsy, were reported as cells/mm². The definition of the invasive margin was a region of 360 µm width on the frontier between malignant cells and peritumoral stroma, which was identical to that reported by Pagès et al.10 (link)

Zhang B., Huang J., Tang J., Hu S., Luo S., Luo Z., Zhou F., Tan S., Ying J., Chang Q., Zhang R., Geng C., Wu D., Gu X, & Liu B. (2021). Intratumoral OH2, an oncolytic herpes simplex virus 2, in patients with advanced solid tumors: a multicenter, phase I/II clinical trial. Journal for Immunotherapy of Cancer, 9(4), e002224.

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