sheep anti gp120 antibody  (Biosynth Carbosynth)

Journal: Nature Cardiovascular Research

Article Title: A bispecific antibody approach for the potential prophylactic treatment of inherited bleeding disorders

doi: 10.1038/s44161-023-00418-4

Figure Lengend Snippet: a , Plasma FVII activity was assessed with a PT-based activity assay in 13 PwGT. Box plot represents the interquartile range. Horizontal line represents the median. Whiskers represent the upper and lower adjacent values. Dotted lines represent the local hospital reference range. b , Plasma FVIIa levels were measured with ELISA in 13 PwGT. Box plot represents the interquartile range. Horizontal line represents the median. Whiskers represent the upper and lower adjacent values. Outliers are indicated as filled black circles. Dotted lines represent the 2.5th and 97.5th percentiles of the FVIIa levels in 50 healthy controls. c , TLT-1 expression was measured with FACS in whole blood of 4 PwGT (green) and 51 healthy controls (gray) in resting platelets or after stimulation with agonists. A labeled IgG4 was used as an isotype control. Box plots represent the interquartile range. Horizontal lines represent the median. Whiskers represent the upper and lower adjacent values. Outliers are shown as filled grey circles. d , Fibrin-dependent platelet pseudoaggregation was measured in four PwGT. Light transmission was monitored for 40 min. Shown are representative traces of light transmission aggregometry in a PwGT (left) and the mean ± s.d. of lag time and the maximum amplitude of the aggregation traces with and without HMB-001 (right, inside the dashed box). Lag time is defined as the time to half-maximal aggregation and is indicated by the vertical dotted lines. Data were analyzed with a two-sided unpaired t test (** P = 0.0019). e , Fibrin-dependent platelet pseudoaggregation was measured in washed platelets from healthy controls supplemented with d -RGDW to obtain GT-like platelets ( n = 3 for each concentration). Aggregation lag time was determined in the presence of FVIIa with or without HMB-001. Data represent mean ± s.d. aggregation lag time at the indicated FVIIa concentration. f , Fibrin-dependent platelet pseudoaggregation was measured in GT-like washed platelets in the presence of 1 nM FVIIa; 0 or 1 nM HMB-001; and 0, 10 or 400 nM sTLT-1. Data are expressed as mean lag time ± s.d. ( n = 3). Data were analyzed with a two-sided unpaired t test (** P = 0.0061). NS, not significant. g , rFVIIa-equivalent activity was measured in PRP from healthy controls ( n = 3) supplemented with d -RGDW to obtain GT-like platelets and a blocking anti-TF antibody to ensure TF independence. FVIIa, FVII and HMB-001 were added at the concentrations obtained by simulating five clinical scenarios (Supplementary Table ). Bars represent the mean rFVIIa-equivalent activity; error bars represent s.d.

Article Snippet: The wells were washed three times, added with 50 μl of the 1:4,000 diluted primary antibody (sheep anti-FVII IgG affinity-purified, Cedarlane) in blocking buffer and incubated for 1 h at RT at 600 r.p.m.

Techniques: Activity Assay, Enzyme-linked Immunosorbent Assay, Expressing, Labeling, Control, Transmission Assay, Concentration Assay, Blocking Assay

Journal: Nature Cardiovascular Research

Article Title: A bispecific antibody approach for the potential prophylactic treatment of inherited bleeding disorders

doi: 10.1038/s44161-023-00418-4

Figure Lengend Snippet: rFVIIa-equivalent activity was measured in PRP from a healthy control (n = 3) supplemented with d -RGDW to obtain GT-like platelets and a blocking anti-TF antibody to ensure TF-independence. PRP was supplemented with rFVIIa (0–75 nM), platelets were stimulated with CRP-XL and PAR-1 AP, and thrombin generation was measured for 240 min. a) For each donor, Endogenous thrombin potential (ETP) was determined in duplicate as a function of rFVIIa concentration. Data represent mean ± s.d. b) ETP was determined in duplicate at 5 predicted clinical scenarios of FVIIa, total FVII(a) and HMB-001 in healthy controls (n = 3). The background plasma concentrations of FVIIa and total FVII(a) were assumed to be 67 pM and 10 nM, respectively. Predicted accumulated levels of total FVII(a) and FVIIa for 5 predicted clinical scenarios (Table ) were reconstituted by adding FVII-S195A and FVIIa after accounting for the background plasma concentration of total FVII(a) and FVIIa respectively. HMB-001 was added as indicated. Data represent mean ± s.d. For each clinical scenario, the corresponding rFVIIa-equivalent activity (nM), as extrapolated from the X-axis of the calibration curve, is indicated next to the horizontal dotted line for the single donor. rFVIIa-equivalent activities for three donors are shown in Fig. .

Article Snippet: The wells were washed three times, added with 50 μl of the 1:4,000 diluted primary antibody (sheep anti-FVII IgG affinity-purified, Cedarlane) in blocking buffer and incubated for 1 h at RT at 600 r.p.m.

Techniques: Activity Assay, Control, Blocking Assay, Concentration Assay