Human recombinant soluble

Recombinant soluble TNF-RI-IgGl fusion protein Etanercept, Enbrel is a chimeric molecule consisting of the extracellular domain of the TNF receptor I (TNF-RI) and the Fc portion of human IgGl. Two Fc domains are bound to each other via disulfide bonds, thereby yielding dimers with two binding sites for the TNF trimer. Etanercept binds with high affinity to extracellular TNF and reduces TNF activity. Etanercept is not effective in Crohn s disease, possibly because it does not lead to destiuction of membrane TNF-a expressing cells. Indications and side effects are similar to those of Infliximab and Adalimumab. 

Mecfianism of Action A recombinant, soluble, long-acting insulin analog that binds to human albumin and provides slow absorption and a prolonged action. Therapeutic Effect Controls glucose levels in diabetic patients. 

Bui, T, C. Faltynek, and R.J. Ho, Differential disposition of soluble and liposome-formulated human recombinant interleukin-7 effects on blood lymphocyte population in guinea pigs. Pharm Res, 1994.11(5) 633-41. 

T.J. Johnston, J.A. Sharma, S.K. Direct interaction of soluble human recombinant T protein with A 1-42 results in t aggregation and hyperpho-sphorylation by t protein kinase IL FEES Lett., 514, 263-268 (2002)... 

In this assay, 100 nM recombinant soluble tissue factor and 2nM recombinant human factor Vila were added to a 96-well assay plate containing 0.4 mM of a selected experimental agent and V-methylsulfonyl-D-Phe-Gly-Arg-p-nitroaniline... 

Hyl was detected in recombinant soluble form of the human CD4 receptor (rCD4) and a related chimeric protein (rCD4-IgG) by amino acid analysis. Tryptic peptides were collected from these proteins and the Hyl sites were identified by N-terminal sequence... 

A recombinant, soluble fusion protein that is a dimer of an extracellular portion of the human TNF receptor and the Fc portion of IgGl (TNFR Fc) binds and neutralizes TNFa and prevents death in animal models of bacteremia and endotoxemia. In patients with septic shock, treatment with the TNFR Fc fusion protein does not reduce mortality, and higher doses appear to be associated with increased mortality. 

PKG and cyclic nucleotide phosphodiesterases (PDEs) (reviewed by Baxter37). A rapid release of natriuretic peptides and induction of their de novo synthesis occurs in myocardial ischemia. Natriuretic peptides may have a cardioprotective role. BNP-32 administration in perfused rat hearts prior to left main coronary artery occlusion and until 30 min of reperfusion resulted in limitation of infarct size in a concentration dependent manner. Furthermore, this effect was abolished by 5-hydroxydecanoate (presumed to be a selective blocker of mitochondrial KATp channels), L-NAME, an inhibitor of NOS and ODQ, a specific soluble guanyl cyclase inhibitor.38 Similarly, human recombinant ANP limited infarct size and reperfusion arrhythmias in a canine model of coronary occlusion and reperfusion.39... 

G22. Gray, P. W., Barrett, K Chantry, D., Turner, M., and Feldmann, M. Cloning of human tumor necrosis factor (TNF) receptor cDNA and expression of recombinant soluble TNF-binding protein. Proc. Natl. Acad. Sci. U.SA. 87, 7380-7384 (1990). 

Table 14.2-4 Predictions of the number of genes in the human genome accessible to protein therapeutics (recombinant soluble proteins and antibodies)...

Figure 4 shows the allometric plots of pharmacokinetic parameters for the recombinant, soluble, and chimeric form of P-selectin glycoprotein ligand-1 (rPSGL-Ig), an antagonist to P-selectin for the treatment of P-selectin-mediated diseases like thrombosis, reperfusion injury, and deep vein thrombosis. Human rPSGL-Ig pharmacokinetic parameters could accurately be predicted on the basis of data from mouse, rat, monkey, and pig using allometric power functions [54]. 

Etanercept is a recombinant form of human tumor necrosis factor receptor.24 Etanercept provides a therapeutic effect by binding to soluble TNF and preventing its binding with TNF receptors. It is administered subcutaneously once or twice weekly noticeable symptomatic improvement is seen in 1 to... 

Adalimumab is a recombinant human IgGl monoclonal antibody specific for human TNF.27 Adalimumab binds to soluble and bound TNF-a. Patients may experience symptomatic relief in as early as 1 week. Adalimumab can be administered in combination with methotrexate or other DMARDs.2... 

TNF is produced and secreted by activated cells within minutes following contact with LPS, reaching peak levels at 90-120 minutes after the admnistration of Escherichia coli endotoxin in human volunteers (M27). Van Deventer et al. (D15) could not detect serum TNF levels during experimental endotoxemia. Even during continuous intravenous administration of recombinant TNF (rTNF), serum TNF rapidly becomes undetectable (M27). It has been proposed that circulating soluble TNF receptors (sTNF-Rs) may be an important down-regulating mechanism (G10). 

U1. Uchiba, M., Okajima, K Murakami, K., Nawa, K., Okabe, H and Takatsuki, K., Recombinant human soluble thrombomodulin reduces endotoxin-induced pulmonary vascular injury via protein C activation in rats. Thromb. Haemost. 74, 1265-1270 (1995). 

The first hurdle encountered during the development of alfalfa as a recombinant protein production system was the relative inefficiency of the available expression cassettes. A study in which a tomato proteinase inhibitor I transgene was expressed in tobacco and alfalfa under the control of the cauliflower mosaic virus (CaMV) 35S promoter showed that 3-4 times more protein accumulated in tobacco leaves compared to alfalfa leaves [5]. Despite the low efficiency of the CaMV 35S promoter in alfalfa, bio-pharmaceutical production using this system has been reported in the scientific literature. Such reports include expression of the foot and mouth disease virus antigen [6], an enzyme to improve phosphorus utilization [7] and the anti-human IgG C5-1 [8]. In this last work, the C5-1 antibody accumulated to 1% total soluble protein [8]. 

High temperatures can break native S-S bonds and form new S-S bonds which can lock the protein into a denatured eonfiguration [89]. Low pH, sodium dodecyl sulfate. Tween 80, chaotropie salts, and exogenous proteins have been used to protect proteins from thermal inaetivation [90]. Ethylene glycol at 30-50% was used to protect the antiviral activity of P-interferon preparations [91]. Human serum albumin was used in recombinant human interferon-Psei-n which resulted in increased thermal stability [62]. Water-soluble polysaeeharides sueh as dextrans and amylose [92], as well as point-specific (site-directed) mutagenesis [93] have also been used to increase thermal stability of therapeutie proteins and peptides. 

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