To remodel the pre-existing FVIII inhibitor development in hemophilia A mice, FVIII–/– mice were treated with rhFVIII (Advate, Baxter, Westlake Village, CA, United States) at a dose of 100 IU/kg, once a week for 4 weeks. The titer of anti-human FVIII inhibitor was measured by using the Bethesda assay, as previously described, using a STart 4 Coagulation Analyzer (Diagnostica Stago, Parsippany, NJ, United States) (29 (link)).
Advate
Manufactured by Baxter
Sourced in United States
Advate is a recombinant coagulation factor VIII (rFVIII) product used for the treatment and prevention of bleeding episodes in patients with hemophilia A. It is a purified protein that helps the blood to clot.
Automatically generated - may contain errors
Lab products found in correlation
Start4 coagulation analyzer, by Diagnostica Stago (1 mentions)
Human albumin, by Baxter (1 mentions)
5 alpha competent e coli cells, by New England Biolabs (1 mentions)
Plasmid maxi kit, by Qiagen (1 mentions)
Porcine thyroglobulin, by Merck Group (1 mentions)
Naocl, by Merck Group (1 mentions)
Lps from e coli o55 b5, by Merck Group (1 mentions)
Feso4, by Merck Group (1 mentions)
Nunc maxisorp, by Thermo Fisher Scientific (1 mentions)
Humate p, by CSL Behring (1 mentions)
Xyntha, by Pfizer (1 mentions)
Kovaltry, by Bayer (1 mentions)
Kogenate fs, by Bayer (1 mentions)
6 protocols using advate
1
FVIII Inhibitor Development in Hemophilia A Mice
2
Platelet-Rich Fibrin for Bone Regeneration
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BMDC transplantation included the production and application of platelet gel (platelet-rich fibrin [PRF]) in order to apply growth factors and a fibrin clot to improve the implantation of the biomaterial and promote the regeneration. Clotting factors were infused 30 minutes before drawing the venous blood: ADVATE (Baxter, Westlake Village, CA) for four patients, AIMAFIX (Kendrion, Lucca, Italy) for patient 3, type B hemophilia. A total of 120 mL of venous blood was drawn yielding 6 mL of PRF utilizing the Vivostat system (Vivolution, Birkeroed, Denmark). The first step of the Vivostat procedure requires 60 mL of plasma and batroxobin to be warmed for 10 minutes at 37°C. This allows fibrinogen to release fibrin peptide A. The resultant fibrin I polymer becomes soluble in acid without the activation of Factor XIII, which was then isolated by centrifugation and added to sodium acetate buffer at the concentration of 0.2 mole/L at pH 4. In presence of Ca2+ at neutral pH (later applied in the operating room), endogenous prothrombin was converted to thrombin, cleaving the fibrinopeptide B from fibrin I to fibrin II. This yields fibrin-platelet concentrations that are 7 to 10 times greater than the baseline values. So PRF is a product capable of providing both hemostasis and tissue regeneration.
3
Virus Inactivation Process Matrices
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As a model matrix for plasma‐derived products, Fraction II was used, that is, the process intermediate obtained during immunoglobulin production at the stage before virus inactivation/reduction steps.22 The starting material had a pH of 5.2. Immediately before virus inactivation runs, the material was filtered through a 0.2 µm filter, and the protein concentration was adjusted to 28.9 AU280‐320/cm with 30 mM NaCl. As a model matrix for recombinant protein production, human albumin (25%, Baxter AG) was diluted to 0.6 mg/ml using a buffer that contained 396 mM NaCl, 20 mM MES acid monohydrate, 10 mM CaCl2 dihydrate, and 0.099% (v/v) PS80. This matrix is representative for an intermediate of the production process of recombinant factor VIII (ADVATE; Baxter AG). The starting material had a pH of 6.4 and was filtered through a 0.2 µm filter immediately before virus inactivation runs were performed.
4
Recombinant Antibody Production Protocol
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The coding sequences of the PG9, PGT128, VRC01 bN-mAbs were obtained from published sequences (available from the NIH AIDS Reagent Program, Germantown, MD) and used to created synthetic genes for stable or transient expression of antibodies in CHO cells. Antibodies were purified using Protein G chromatography. These cell lines have been deposited in the NIH AIDS Reagent Program. The 10–1074 bN-mAb was acquired from the NIH AIDS Reagent Program. The 34.1 mAb to the Herpes Simplex Virus glycoprotein D (gD) was produced in-house [39 (link)]. The human 447-52D mAb was contributed by Dr. Susan Zolla-Pazner (Mount Sinai, New York City, NY). Recombinant rgp120s were cloned with an N-terminal gD tag into a pCF vector [39 (link)]. Plasmids were transformed in 5-alpha competent E. coli cells (New England Biolabs, Ipswich, MA), purified using Plasmid Maxi Kit (Qiagen, Hilden, Germany) and electroporated into CHO cells as described below. Recombinant human factor VIII consisted of Advate (Baxter Healthcare Corporation, Deerfield, Illinois) [40 (link)], and was acquired by the laboratory of Dr. Peter Lollar (Emory University, Atlanta, GA).
5
Oxidation Impacts on Therapeutic Antibodies
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Ninety six-well polystyrene plates NUNC MaxiSorp™ (Thermo Fisher Scientific) were coated for 90 min at room temperature with human recombinant factor VIII (Advate, Baxter), human plasma-derived C3 (CompTech), porcine thyroglobulin (Sigma-Aldrich), bovine histone 3 (Sigma-Aldrich), recombinant LysM polypeptide of AtlA from Enterococcus faecalis and LPS from E. coli O55:B5 (Sigma-Aldrich). The coating was performed with protein diluted to 2 μg/mL (Factor VIII and C3) or 10 μg/mL (thyroglobulin, histone 3, LysM and LPS) in PBS. Afterwards, the plates were blocked by incubation for 1 h at room temperature with a solution of 0.25% Tween 20 in PBS. Rituximab and Trastuzumab (both stock solutions in PBS) were first diluted to 1 mg/mL (6.7 μM) in PBS and left intact or exposed for 10 min at room temperature to 500 μM FeSO4 or to 200 μM NaOCl (Sigma-Aldrich). After native substances and those treated with pro-oxidative Abs were serially diluted from 3.35 to 0.003 μM and incubated for 1 h at room temperature. The following steps of the ELISA are identical with those described above.
6
Comparative Study of FVIII and VWF-FVIII Products
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