Journal: mAbs

Article Title: Development and pharmacokinetic assessment of a fully canine anti-PD-1 monoclonal antibody for comparative translational research in dogs with spontaneous tumors

doi: 10.1080/19420862.2023.2287250

Figure Lengend Snippet: A. Primary structures of lead canine anti-cPD-1 antibodies. A. (top) heavy chain alignments of P4B1, P3C6 and its variants along with canine germline gene IGHV3-38*01. (bottom) light chain alignments of P4B1 and P3C6. Framework and CDR regions use IMGT nomenclature. B. Evaluation of full-length IgG D (P3C6mut3.1 and mut3.2) canine anti-cPD-1 binding to cell surface-expressed cPD-1 by flow cytometry. Canine IgG D -HA tagged clones against cPD-1 or the irrelevant MERS antigen (negative control) were used to stain KTδ32 and KTδ32.cPD-1 cells. Bound IgG D molecules were detected using an anti-HA antibody. Plots are gated on live, 7AAD- cells.

Article Snippet: The VH and VL chains of selected scFvs were cloned into separate expression plasmids engineered to express either canine constant light kappa (InvivoGen, pfuse2-dclk), constant light lambda (InvivoGen, pfuse2-dcll), or constant IgG D (analogous to human IgG4) heavy-chain domain (InvivoGen, pfuse-dchg4).

Techniques: Binding Assay, Flow Cytometry, Clone Assay, Negative Control, Staining

Journal: mAbs

Article Title: Development and pharmacokinetic assessment of a fully canine anti-PD-1 monoclonal antibody for comparative translational research in dogs with spontaneous tumors

doi: 10.1080/19420862.2023.2287250

Figure Lengend Snippet: Full-length IgG D P3C6mut3.1 reverses the effects of PD-L1 inhibition on canine CAR-T cells. A. K562 cells were engineered to express canine CD20 (K562-cCD20) and canine PD-L1 (K562-cCD20-cPD-L1) and cell surface expression was confirmed by flow cytometry. B. Canine CD20 CAR-T cells from one dog were labeled with cell trace violet (CTV) and co-cultured at an E:T ratio of 1:1 with either K562-cCD20 or K562-cCD20-cPD-L1 in the presence of either anti-MERS or P3C6mut3.1 IgG D . After 72 hours of culture, proliferation of CD8 + CAR-T cells was assessed by flow cytometry. Plots are gated on live CD5 + CD8 + CAR + cells. C. Canine CD20 CAR-T cells from one dog were co-cultured at an E:T ratio of 1:1 with the same target cells as in A. Expression of CD107b was determined after 4 hours of co-culture. Plots are gated on live CD5 + CD8 + CAR + cells.

Article Snippet: The VH and VL chains of selected scFvs were cloned into separate expression plasmids engineered to express either canine constant light kappa (InvivoGen, pfuse2-dclk), constant light lambda (InvivoGen, pfuse2-dcll), or constant IgG D (analogous to human IgG4) heavy-chain domain (InvivoGen, pfuse-dchg4).

Techniques: Inhibition, Expressing, Flow Cytometry, Labeling, Cell Culture, Co-Culture Assay

Journal: mAbs

Article Title: Development and pharmacokinetic assessment of a fully canine anti-PD-1 monoclonal antibody for comparative translational research in dogs with spontaneous tumors

doi: 10.1080/19420862.2023.2287250

Figure Lengend Snippet: Pharmacokinetics of full-length IgG D/B P3C6mut3.1 in healthy dogs. Healthy dogs were administered P3C6mut3.1 IgG D/B intravenously at either 2 mg/kg ( n = 2) or 10 mg/kg ( n = 2) on Day 0 and Day 21. Blood samples were taken at the indicated time points and analyzed for the presence of P3C6mut3.1 IgG D/B in the serum using a custom Meso Scale Discovery immunoassay. A. Serum pharmacokinetics of P3C6mut3.1 IgG D/B with data represented as mean ± SD. B. Dose-normalized concentration vs. time profiles for P3C6mut3.1 IgG D/B .

Article Snippet: The VH and VL chains of selected scFvs were cloned into separate expression plasmids engineered to express either canine constant light kappa (InvivoGen, pfuse2-dclk), constant light lambda (InvivoGen, pfuse2-dcll), or constant IgG D (analogous to human IgG4) heavy-chain domain (InvivoGen, pfuse-dchg4).

Techniques: Concentration Assay

Journal: mAbs

Article Title: Development and pharmacokinetic assessment of a fully canine anti-PD-1 monoclonal antibody for comparative translational research in dogs with spontaneous tumors

doi: 10.1080/19420862.2023.2287250

Figure Lengend Snippet: A non-compartmental analysis of full-length P3C6mut3.1 IgG D/B PK parameters following the first dose. C max : maximum observed serum concentration, AUC last : area under the serum concentration vs. time curve from time 0 to 21 days, AUC inf : area under the serum concentration vs. time curve extrapolated to time infinity, t 1/2 : terminal log-linear half-life, CL: clearance, V ss : volume of distribution.

Article Snippet: The VH and VL chains of selected scFvs were cloned into separate expression plasmids engineered to express either canine constant light kappa (InvivoGen, pfuse2-dclk), constant light lambda (InvivoGen, pfuse2-dcll), or constant IgG D (analogous to human IgG4) heavy-chain domain (InvivoGen, pfuse-dchg4).

Techniques: Concentration Assay