Determination of biologically active forms of proteolytic enzymes

   
   

A process is provided for the assessment of an active proteolytic enzyme in a blood or another biological fluid sample possibly comprising a complex of said proteolytic enzyme and .alpha..sub.2 -macroglobulin, wherein said sample is contacted with a substrate comprising a molecule of sufficient size coupled to a signal-substrate, said signal-substrate comprising a detectable leaving group, wherein said substrate is hydrolysed by said proteolytic enzyme but not by said complex. The proteolytic enzyme is preferably selected from the group consisting of thrombin, activated clotting factor, activated fibrinolytic factor, and activated component of the complement system. Of these, thrombin is most preferred. The molecules of sufficient size are preferably water soluble and selected from the group consisting of inert protein, preferably ovalbumin, polysaccharide, and synthetic polymer. The size of these molecules is such that they will not fit into the cavity of the .alpha..sub.2 M molecules.

Um processo é fornecido para a avaliação de um enzyme proteolytic ativo em um sangue ou em uma outra amostra fluida biológica que compreendem possivelmente um complexo de enzyme e de alpha..sub.2 proteolytic ditos - macroglobulin, wherein a amostra dita é contatada com uma carcaça que compreende uma molécula do tamanho suficiente acoplada a uma sinal-carcaça, sinal-carcaça dita que compreende um grupo saindo detectável, wherein a carcaça dita hydrolysed por enzyme proteolytic dito mas não por complexo dito. O enzyme proteolytic é selecionado preferivelmente do thrombin consistindo do grupo, do fator clotting ativado, do fator fibrinolítico ativado, e do componente ativado do sistema do complemento. Destes, o thrombin é o mais preferido. As moléculas do tamanho suficiente são preferivelmente água soluble e selecionada do grupo que consiste na proteína inerte, preferivelmente ovalbumin, polysaccharide, e polímero sintético. O tamanho destas moléculas é tal que não caberão na cavidade das moléculas do alpha..sub.2 M.

 
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> High yield S-nitrosylation process

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