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Be0032

Manufactured by BioXCell

The BE0032 is a laboratory centrifuge designed for a variety of applications. It features a compact and durable construction, making it suitable for use in various research and testing environments. The centrifuge is capable of reaching high speeds and can accommodate different types of sample containers, enabling efficient separation and processing of materials. Further details on the specific capabilities and intended uses of this product are not available.

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6 protocols using be0032

1

Anti-CTLA-4 and Anti-PD-L1 Therapy in Murine Tumors

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Studies of anti–CTLA-4 (BE0032, Bio X Cell) and anti–PD-L1 (BE0101, Bio X Cell) therapy in mice bearing 4T1 and LLC tumors, as well as studies of scAAV monotherapy and combination treatment, are consistent with the descriptions above. The administration methods of each drug are shown in the respective flow charts in the figures.
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2

Tumor Inhibition by PEGylated Kynureninase

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Example 21

The PEGylated Pseudomonas fluorescence kynureninase (PEG-Pf-KYNU) was evaluated in the B16 melanoma mouse model in a side-by-side comparison with the anti-PD1 (clone RMP1-14, BioXCell #BE0146) or anti-CTLA-4 (clone UC10-4F10-11, BioXCell #BE0032) immune checkpoint inhibitor antibodies. Fifty thousand B16 cells were implanted in the flank of C57BL/6J mice (Day 0, n=8 mice each group). Once palpable tumors developed (Day 10), the animals were treated with either 250 μg anti-PD1, 100 μg anti-CTLA-4 (200 μg 1st dose as per Holmgaard et al. (2013)), or 500 μg of PEG-Pf-KYNU at the times shown (FIG. 8). Heat-inactivated PEG-Pf-KYNU was used as a control. Administration of PEG-Pf-KYNU resulted in significant tumor growth retardation and extended survival in a manner indistinguishable from that observed with the anti-PD1 or anti-CTLA-4 checkpoint inhibitor antibodies (FIG. 8) for PEG-Pf-KYNU vs. inactivated enzyme or PBS only.

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3

PEG-Pf-KYNU Inhibits Melanoma Growth

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Example 21

The PEGylated Pseudomonas fluorescence kynureninase (PEG-Pf-KYNU) was evaluated in the B16 melanoma mouse model in a side-by-side comparison with the anti-PD1 (clone RMP1-14, BioXCell #BE0146) or anti-CTLA-4 (clone UC10-4F10-11, BioXCell #BE0032) immune checkpoint inhibitor antibodies. Fifty thousand B16 cells were implanted in the flank of C57BL/6J mice (Day 0, n=8 mice each group). Once palpable tumors developed (Day 10), the animals were treated with either 250 μg anti-PD1, 100 μg anti-CTLA-4 (200 μg 1st dose as per Holmgaard et al. (2013)), or 500 μg of PEG-Pf-KYNU at the times shown (FIG. 8). Heat-inactivated PEG-Pf-KYNU was used as a control. Administration of PEG-Pf-KYNU resulted in significant tumor growth retardation and extended survival in a manner indistinguishable from that observed with the anti-PD1 or anti-CTLA-4 checkpoint inhibitor antibodies (FIG. 8) for PEG-Pf-KYNU vs. inactivated enzyme or PBS only.

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4

Comparison of PEG-Pf-KYNU and Checkpoint Inhibitors

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Example 21

The PEGylated Pseudomonas fluorescence kynureninase (PEG-Pf-KYNU) was evaluated in the B16 melanoma mouse model in a side-by-side comparison with the anti-PD1 (clone RMP1-14, BioXCell # BE0146) or anti-CTLA-4 (clone UC10-4F10-11, BioXCell # BE0032) immune checkpoint inhibitor antibodies. Fifty thousand B16 cells were implanted in the flank of C57BL/6J mice (Day 0, n=8 mice each group). Once palpable tumors developed (Day 10), the animals were treated with either 250 μg anti-PD1, 100 μg anti-CTLA-4 (200 μg 1st dose as per Holmgaard et al. (2013)), or 500 μg of PEG-Pf-KYNU at the times shown (FIG. 8). Heat-inactivated PEG-Pf-KYNU was used as a control. Administration of PEG-Pf-KYNU resulted in significant tumor growth retardation and extended survival in a manner indistinguishable from that observed with the anti-PD1 or anti-CTLA-4 checkpoint inhibitor antibodies (FIG. 8) for PEG-Pf-KYNU vs. inactivated enzyme or PBS only.

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5

PEGylated Kynureninase Inhibits Melanoma

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Example 21

The PEGylated Pseudomonas fluorescence kynureninase (PEG-Pf-KYNU) was evaluated in the B16 melanoma mouse model in a side-by-side comparison with the anti-PD1 (clone RMP1-14, BioXCell #BE0146) or anti-CTLA-4 (clone UC10-4F10-11, BioXCell #BE0032) immune checkpoint inhibitor antibodies. Fifty thousand B16 cells were implanted in the flank of C57BL/6J mice (Day 0, n=8 mice each group). Once palpable tumors developed (Day 10), the animals were treated with either 250 μg anti-PD1, 100 μg anti-CTLA-4 (200 μg 1st dose as per Holmgaard et al. (2013)), or 500 μg of PEG-Pf-KYNU at the times shown (FIG. 8). Heat-inactivated PEG-Pf-KYNU was used as a control. Administration of PEG-Pf-KYNU resulted in significant tumor growth retardation and extended survival in a manner indistinguishable from that observed with the anti-PD1 or anti-CTLA-4 checkpoint inhibitor antibodies (FIG. 8) for PEG-Pf-KYNU vs. inactivated enzyme or PBS only.

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6

Immunomodulation in Paracoccidioidomycosis

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Three weeks after P. brasiliensis infection, groups of mice received three times a week intraperitoneal (ip.) injections of 200 μg contained in 100 μl of mAb α-CTLA-4 (BioXCell Cat # BE0032 clone UC10F10-11, hamster IgG)), α-PD-1 (CD279, BioXCell, Cat # BE0146, clone RMP1-14, rat IgG2a), control monoclonal IgG ((BioXCell, Cat # BE0089, clone 2A3, rat IgG2a), or 100 μl of PBS (Phosphate Buffered Saline). All mAbs contained <2EU/mg of endotoxin (<0.002EU/μg) as determined by LAL gel clotting assay. The dose of α-CTLA-4 and α-PD-1 mAbs corresponds to approximately 10 μg/kg of body weight. Treatment was started in the third week post-infection because in this period the adaptive immunity and the expansion of its regulatory mechanisms, such as the expression of CTLA-4 and PD-1, are already established in infected C57BL/6 mice (23 (link)–26 (link)). The treatment was maintained for five weeks. Animals were sacrificed eight weeks after infection, and the immune response and disease severity were analyzed by various methods. The experiments were repeated twice.
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