Anti pd l1 antibody

Manufactured by BioXCell

Sourced in United States, Lebanon

The Anti-PD-L1 antibody is a laboratory reagent designed for research purposes. It is a monoclonal antibody that specifically binds to the programmed death-ligand 1 (PD-L1) protein. PD-L1 is a molecule involved in the regulation of the immune system. The antibody can be used in various research applications to study the interaction between PD-L1 and its receptor, PD-1, and their role in immune responses.

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20 protocols using anti pd l1 antibody

Lung Metastasis Modeling and Immunotherapy

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Eight-week-old mice were injected intravenously with 5 × 10⁵ KP cells^{22 (link)}, KP–GFP cells or B16-BFP/OVA cells. All cell lines tested negative for mycoplasma and were authenticated by phenotyping their potential for generating tumours in mice. Lungs and lymph nodes were analysed on day 28 (KP or KP–GFP) or on day 22 (B16-BFP/OVA), except when otherwise indicated. When indicated, mice were injected intraperitoneally (i.p.) with 25 μg anti-IL-4 antibody (BioXcell, clone 11B11) on days 21, 23 and 26; with 100 μg CD40 agonistic antibody (BioXcell clone FGK4.5/ FGK45) on day 27; with 200 μg poly(I:C) high-molecular-weight RNA (InvivoGen) on day 27; or with 200 μg anti-PD-L1 antibody (BioXcell, clone 10F.9G2) on days 15, 18, 21, 24 and 27.
To quantify tumours, we stained slides of paraffin-embedded left lung lobes with haematoxylin/eosin; we scanned slides using an Olympus digital scanner and analysed them using Panoramic Viewer software.

Maier B., Leader A.M., Chen S.T., Tung N., Chang C., LeBerichel J., Chudnovskiy A., Maskey S., Walker L., Finnigan J.P., Kirkling M.E., Reizis B., Ghosh S., D’Amore N.R., Bhardwaj N., Rothlin C.V., Wolf A., Flores R., Marron T., Rahman A.H., Kenigsberg E., Brown B.D, & Merad M. (2020). A conserved dendritic-cell regulatory program limits antitumour immunity. Nature, 580(7802), 257-262.

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Combination Immunotherapy for Glioblastoma

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Anti-PDL1 antibody, anti-CD4 depletion antibody, and anti-CD40 agonist were purchased from BioXCell. For scRNA-seq experiments, when indicated, 200 mg CD4 depleting antibody was administered intraperitoneally on day 5 after tumor implantation, and 200 mg of anti-PDL1 was administered intraperitoneally on day 11 after tumor implantation. TIL were harvested on day 14 post implantation. For survival experiments, anti-PDL1 and CD40 agonist were dosed as follows: GL261—CD40 agonist on day 9, anti-PDL1 on days 11, 15, and 19; and CT2A—CD40 agonist on day 7, anti-PDL1 on days 9, 12, 16, and 21 for CT2A. CD4 depletion antibody was administered on day 5 and weekly thereafter. Mortality was recorded or mice were euthanized on reaching predetermined humane endpoints after daily monitoring for neurological deficits, signs of suffering, or impaired function. Kaplan-Meier survival analyses were conducted in GraphPad Prism using Mantel-Cox test. All mice were included for scRNA-seq and survival analyses.

Khan S.M., Desai R., Coxon A., Livingstone A., Dunn G.P., Petti A, & Johanns T.M. (2022). Impact of CD4 T cells on intratumoral CD8 T-cell exhaustion and responsiveness to PD-1 blockade therapy in mouse brain tumors. Journal for Immunotherapy of Cancer, 10(12), e005293.

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Murine Tumor Implantation and Anti-PD-L1 Therapy

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MC38 cells and B16F10 cells were purchased from ATCC and cultured in complete DMEM-10 medium (Gibco) supplied with 10% FBS (Gibco), 1% penicillin/streptomycin (Gibco), and 1% L-glutamine (Gibco). C57BL/6 mice were subcutaneously implanted with 2 × 10⁵ MC38 cells or 2 × 10⁵ B16F10 cells at the anterolateral aspect of the left thigh. Tumor volumes were measured with a caliper and calculated according to the formula: ((length × width²)/2). Tumor-engrafted mice were sacrificed at indicated timepoints on the premise of the humane endpoints (tumor volume not exceeding 2,000 mm³). For anti-PD-L1 antibody treatment experiments, mice were intraperitoneally injected with 200 μg anti-PD-L1 antibody (BioXcell, clone 10F.9G2) at indicated time points.

Chen X., Lin Y., Yue S., Yang Y., Yang X., He J., Gao L., Li Z., Hu L., Tang J., Wang Y., Tian Q., Hao Y., Xu L., Huang Q., Cao Y, & Ye L. (2023). PD-1/PD-L1 blockade restores tumor-induced COVID-19 vaccine bluntness. Vaccine.

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PD-L1 Blockade and miRNA Modulation

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Surface PD-L1 on NB cells was blocked by treatment with control immunoglobulin (Ig) G or anti-PD-L1 antibody (15 μg/mL, BioXCell, Lebanon, NH) for 24 h followed by transfection with miR-15a and miR-15b oligonucleotides for an additional 24 h. After wash out the unbound antibody, these cells were used for coculture experiments with CD8⁺T and NK cells.

Pathania A.S., Prathipati P., Olwenyi O.A., Chava S., Smith O.V., Gupta S.C., Chaturvedi N.K., Byrareddy S.N., Coulter D.W, & Challagundla K.B. (2022). miR-15a and miR-15b modulate natural killer and CD8+T-cell activation and anti-tumor immune response by targeting PD-L1 in neuroblastoma. Molecular Therapy Oncolytics, 25, 308-329.

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Mouse Tumor Model for Immunotherapy

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All animal experiments were approved by the Animal Care and Use Committee of the Institute of Materia Medica, Chinese Academy of Medical Sciences, and Peking Union Medical College (Beijing, China). Five-to six-week-old female hPD-1 C57BL/6 mice were obtained from the Nanjing Biomedical Research Institute of Nanjing University (Nanjing, China). hPD-L1 MC38 cell lines were cultured and harvested in saline. A total of 5 × 10⁵ cells were injected subcutaneously into the right flank of the mice. When the average tumor volume reached 100–200 mm³, the mice were randomly enrolled in the control and experimental groups (n = 7) and treatment was started on Day 1. For the control group, 0.5% (w/v) sodium carboxymethyl cellulose Na (CMCNa) was administered orally every day. YPD-30, a prodrug for YPD-29B with significantly higher bioavailability than YPD-29B, was dissolved in 0.5% CMCNa for oral treatment every day and anti-PD-L1 antibody (Bio X Cell, Lebanon, NH, USA) was dissolved in saline for intraperitoneal treatment twice a week. Tumor volumes were measured twice a week. At the end of the experiment, the mice were sacrificed and the tumors were collected, weighted, and analyzed. The tumor volume was calculated as Eq. (3): in which L is the maximum tumor length and W is the maximum tum-or width. Tumor growth inhibition (TGI) was calculated as Eq. (4):

Lai F., Ji M., Huang L., Wang Y., Xue N., Du T., Dong K., Yao X., Jin J., Feng Z, & Chen X. (2022). YPD-30, a prodrug of YPD-29B, is an oral small-molecule inhibitor targeting PD-L1 for the treatment of human cancer. Acta Pharmaceutica Sinica. B, 12(6), 2845-2858.

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Antibody Conjugation and Characterization of Anti-PD-L1 TKNP

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DPEMA used as a surfactant during TKNP synthesis provides an active site for the conjugation of anti‐PD‐L1 antibody.¹⁵ To conjugate anti‐PD‐L1 antibody (Bio X Cell, Lebanon, NH), TKNP (1 mg) was incubated with different concentrations of anti‐PD‐L1 antibody (30, 60, 90, 120, and 150 μg/ml) suspended in NaHCO₃ buffer (pH 8.5). Following incubation for 1 h, repeated purification was conducted at 15,000 × g for 10 min at 4°C. To determine the conjugated amount of antibody on TKNP surface, Pierce BCA protein assay kit (Thermo Fisher Scientific) was used. CE were calculated using the formulae:

CE (%) = (\frac{Amount of anti - PD - L 1 antibody conjugated to TKNP}{Initial amount of anti - PD - L 1 antibody used}) \times 100

Similarly, DLS was used to monitor the mean hydrodynamic diameter, PDI, and zeta potential of the anti‐PD‐L1‐TKNP.

Banstola A., Pandit M., Duwa R., Chang J., Jeong J, & Yook S. (2022). Reactive oxygen species‐responsive dual‐targeted nanosystem promoted immunogenic cell death against breast cancer. Bioengineering & Translational Medicine, 8(5), e10379.

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Synthesis and Evaluation of YSK12-C4 Nanoparticles

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YSK12-C4 was synthesized as previously described [14 (link)]. The commercially available reagents used are as follows: Cholesterol (Sigma-Aldrich, St. Louis, MO, USA); 1,2-Dimirystoyl-sn-glycerol methoxyethyleneglycol 2000 ether (DMG-PEG2k) (NOF Corporation, Tokyo, Japan); Cyclic di GMP (c-di-GMP) (Cayman chemical, Ann Arbor, MI, USA); anti-programmed cell death ligand 1 (anti-PD-L1) antibody (clone: 10F.9G2), anti-CTLA-4 antibody (clone: 9H10), and anti-lymphocyte activation gene 3 (anti-LAG-3) antibody (clone: C9B7W) (Bio X Cell, West Lebanon, NH, USA).
Renca cells were purchased from the American Type Culture Collection (Manassas, VA, USA) and were cultured with RPMI1640 medium (high glucose) containing 10% fetal bovine serum (FBS), 100 units/mL of penicillin/streptomycin (P/S), and 1 mM of sodium pyruvate.
Female BALB/c mice (6–8 weeks old) (Japan SLC Inc., Shizuoka, Japan) were housed under specific pathogen-free (SPF) conditions. The animal experiments described herein were approved by the Animal Committee of Hokkaido University (approval number: 20-0176). All methods were conducted based on the guidelines set by Hokkaido University and the guidelines established for Animal Research: Reporting of In Vivo Experiments (ARRIVE).

Nakamura T., Sasaki S., Sato Y, & Harashima H. (2023). Cancer Immunotherapy with Lipid Nanoparticles Loaded with a Stimulator of Interferon Genes Agonist against Renal Tumor Lung Metastasis. Pharmaceutics, 16(1), 31.

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Anti-PD-L1 and Radiation Therapy in Murine Tumor Models

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B16-OVA and AT-84-E7-OVA cells were cultured and injected subcutaneously in the right flank with 1.5 × 10^5 cells (B16-OVA) or 5 × 10^5 cells (AT-84-E7-OVA) on day 0. The injection sites were monitored until a tumor was palpable. The tumor was then measured using electronic calipers every 3 days and tumor volume was calculated using (l x ŵ2)/2. Anti-PD-L1 therapy and RT was started as soon as tumors were palpable. Mice were treated with 200 μg anti-PD-L1 antibody (BioXcell) via intraperitoneal injections every 3 days for a total of 3 injections per mouse. Mice were treated with 12–18 Gy of focal radiation to the tumor site using a JL Shepherd Cs-137 Irradiation (JL Shephard and Associates, San Francisco, CA) a dose rate of 2.53 Gy / min. To direct radiation, customized lead shielding with a jig to immobilize the region receiving radiation were used for each mouse. To deplete B-cells, 250 μg of anti-CD20 antibody (clone SA271G2, Biolegend) was given intravenously through the tail vein following manufacturer instructions 5 days prior to tumor injection.

Kim S.S., Shen S., Miyauchi S., Sanders P.D., Franiak-Pietryga I., Mell L.K., Gutkind J.S., Cohen E.E., Califano J, & Sharabi A.B. (2020). B-cells improve overall survival in HPV-associated squamous cell carcinomas and are activated by radiation and PD-1 blockade.. Clinical cancer research : an official journal of the American Association for Cancer Research, 26(13), 3345-3359.

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Tumor Growth Inhibition by Compound 15a in Mice

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A total of 2 × 10⁵ 4T1 cells were subcutaneously inoculated into female BALB/c wild-type mice aged 6–8 weeks, which were obtained from Guangdong Pharmaceutical Company. Once the tumour volume reached 80–100 mm³, the mice were randomly assigned to two groups, with five mice in each group. The groups were administered either a vehicle (10% DMSO, 45% PEG300, and 45% 1× PBS) or 15a (dissolved in vehicle) intravenously once daily. Tumour size was measured using vernier calipers every three days. The mean tumour volume for each group was calculated using the formula (length × width × width)/2 and expressed in cubic millimeters. The animals were killed on day 21, and the removed tumour tissue was weighed and photographed. For in vivo combination therapy, mice were treated with vehicle or 15a alone injected on once every three days or in combination with anti-PD-L1 antibody (BioXcell, 100 µg injected on days 7, 10, 13). The experiments were conducted in accordance with the regulations of the Agency’s Animal Management and Use Committee and were approved by the Committee.

Huang W., Zhu Q., Shi Z., Tu Y., Li Q., Zheng W., Yuan Z., Li L., Zu X., Hao Y., Chu B, & Jiang Y. (2024). Dual inhibitors of DNMT and HDAC induce viral mimicry to induce antitumour immunity in breast cancer. Cell Death Discovery, 10, 143.

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Lentiviral Vector Vaccination and LCMV Infection in Mice

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For prophylactic vaccination, C57BL/6 (Taconic Biosciences) or SAMHD1-KO (provided by Axel Roers, Technische Universität Dresden, Dresden, Germany; ref. 52 (link)) mice were injected i.p. or i.v. with 2.5 × 10⁶ IU lentiviral vector. Seven days postvaccination, the mice were injected with 2.0 × 10⁵ PFU LCMV Armstrong (provided by Dirk Homann, Mount Sinai, New York, New York, USA). Four days postinfection, splenic virus load was measured by real-time qRT-PCR. For chronic LCMV infection, SAMHD1-KO mice were challenged by i.v. injection of 5 × 10⁶ PFU LCMV clone 13. Two weeks postinfection, the mice were injected with 3 × 10⁶ IU of lentivirus. Serum was collected weekly and virus load was measured. One week postinjection, the transduced BMDCs were injected i.p. or i.v. Where indicated, the mice were injected 3 times every third day with 30 to 50 μg anti–PD-L1 antibody (Bio X Cell), clone 10F.9G2, BP0101).

Tada T., Norton T.D., Leibowitz R, & Landau N.R. (2022). Directly injected lentiviral vector–based T cell vaccine protects mice against acute and chronic viral infection. JCI Insight, 7(18), e161598.

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