Recombinant human tissue plasminogen activator

Activase (Alteplase) [CHO-expressed glycoprotein-recombinant human tissue plasminogen activator] Genentech Lyophilized (50 mg single-dose vial 100 mg single dose vial) 50 and 100 mg doses to be reconstituted with 50 or lOOmL of WFI, respectively Per 50 mg vial 1.7 g L-arginine 0.5 g phosphoric acid < 4 mg polysorbate 80 (under vacuum) Per 100 mg vial 3.5 g L-arginine 1 g phosphoric acid <11 mg polysorbate 80 (without vacuum) IV... 

Zhang L, Zhang ZG, Liu X, Hozeska A, StagUano N (2006) Treatment of embolic stroke in rats with bortezomib and recombinant human tissue plasminogen activator. Thromb Haemost 95 166-173... 

During detailed characterization of a tryptic map of recombinant human tissue plasminogen activator (rtPA), a minor peak was resolved whose mass was not consistent with the expected set of tryptic peptides (L. Keyt and S.-L. Wu, unpublished observation). N-terminal sequence analysis of this fraction showed that it had a sequence containing residues 276-296 of rtPA with Lys-277 present as Hyl. We developed a modified amino acid analysis program capable of detecting hydroxylysine at levels down to 0.05 residues/mol to determine the distribution of Hyl in rtPA as well as in some other proteins derived from mammalian cells. 

Zhang L, Zhang ZG, Zhang RL, Lu M, Krams M, Chopp M (2003) Effects of a selective CDl lb/ CD 18 antagonist and recombinant human tissue plasminogen activator treatment alone and in combination in a rat embolic model of stroke. Stroke 34 1790-1795... 

USA alteplase antithrombotic Activase Actilyse, Activacin, Cathflo Activase, GRTPA Recombinant human tissue plasminogen activator, glycoform alpha produced in CHO cells Thrombosis... 

Generic Recombinant Human Tissue Plasminogen Activator (tPA) Penicillin Manufacture Novobiocin Manufacture Polymers... 

FIGURE 3 HPLC chromatogram of the tryptic peptides produced from recombinant human tissue plasminogen activator (Mr 60 kDa) with UV monitoring (A) or total ion current (TIC) as measured by the electrospray mass spectrometer (B). Reprinted, with permission, from Ling et al. (1991). Each tryptic peptide is identified by its order in the sequence from 1 to 51. Glycopeptide masses observed are listed in Table I. 

Van Reis et al. [92] reported the scale-up of a HF system for the recovery of human tissue plasminogen activator (t-PA) produced by recombinant CHO cells from the 2.5-m to the 180-m scale. A robust and reproducible process was achieved by combining hnear scale-up principles, control of fluid dynamic parameters and experimentally defined limits of product retention, which meant maintaining channel length, wall shear rate and flux constant. 

The first successful therapeutic protein made in a transgenic system was human tissue plasminogen activator regulated by a milk-directed promoter for accumulation in mouse milk [5]. Human growth hormone, which was one of the first proteins produced using recombinant microbial systems in the early 1980s, became the first human protein expressed in plants (tobacco cells) in 1986 [6]. Since then, over 200 biotherapeutics of diverse origin. 

Papadopoulos SM, Chandler WF, Salamat MS, Topol EJ, Sackellares JC. Recombinant human tissue-type plasminogen activator therapy in acute thromboembolic stroke. J Neurosurg. 1987 67 394-398. 

Yim, K. W., Fractionation of the human recombinant tissue plasminogen activator (rtPA) glycoforms by high-performance capillary zone electrophoresis and capillary isoelectric focusing, /. Chromatogr , 559, 401, 1991. 

Along with the production of insulin, many other medical uses have been achieved for recombinant DNA. This includes the production of erythropoetin, a hormone used to stimulate production of red blood cells in anemic people tissue plasminogen activator, an enzyme that dissolves blood clots in heart attack victims and antihemophilic human factor VIII, used to prevent and control bleeding for hemophiliacs. These three important genetically engineered proteins were all cloned in hamster cell cultures. 

Alteplase Alteplase is a drag that activates human tissue plasminogen (t-PA). It is a glycoprotein of molecular mass 68,000 that is synthesized by vascular endothelial cells. t-PA cells were first isolated from cultured human melanoma cells [56-58], but currently a genetically recombined form of rt-PA is genetically engineered. 

Currently, in both arterial and venous occlusion, newer products such as alteplase which is recombinant human tissue-type plasminogen activator where 10 mg is given as an intitial bolus and a further 90 mg infused over 2 hours are offering alternative regimens. Although costly they have apparent benefit in stroke and acute coronary syndromes. 

Also known as recombinant tissue Plasminogen Activator (rt-PA). It is produced by recombinant DNA technology from human tissue culture. It specifically activates plasminogen bound to the fibrin clot. This minimises the risk of systemic bleeding. 

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