RESUMO
A solid phase, enzyme-linked immunoassay is described for the quantitative determination of the complex of human granulocyte elastase (EC 3.4.21.37) with alpha 1-proteinase inhibitor. The assay employs antibody-coated test tubes and it is suitable for routine use in clinical chemistry laboratories. Data for sample stability and test characteristics are given. A reference range of 20-180 micrograms/l elastase in plasma was determined. The diagnostic significance of granulocyte elastase levels in plasma in inflammatory diseases is discussed.
Assuntos
Proteínas Sanguíneas/análise , Neutrófilos/enzimologia , Elastase Pancreática/sangue , Fenômenos Químicos , Química , Granulócitos/enzimologia , Humanos , Técnicas Imunoenzimáticas , alfa 1-AntitripsinaRESUMO
The purification of plasma proteins by affinity chromatography on triazine dye matrices can be optimised with regard to the triazine dye used as a group-specific ligand. A comparison by electrophoretic and immunological means of the results of affinity chromatography with human plasma on Fractogel TSK-Blue and Fractogel TSK-Red demonstrated that the Procion Red HE-3B-containing gel was able to adsorb more plasma constituents than the Cibacron Blue F3-GA gel. The preparation of alpha 1-proteinase inhibitor obtained after chromatography on Fractogel TSK-Red showed a higher degree of purity and could easily be further purified by ion-exchange chromatography on Fractogel TSK DEAE-650 and gel filtration on Fractogel TSK HW 55, without significant loss of biological activity.
Assuntos
Triazinas , alfa 1-Antitripsina/isolamento & purificação , Cromatografia de Afinidade/métodos , Cromatografia em Gel/métodos , Cromatografia por Troca Iônica/métodos , Eletroforese em Gel de Poliacrilamida/métodos , HumanosRESUMO
Granulocyte elastase in complex with its main inhibitor in plasma, i.e. alpha 1-proteinase inhibitor, was quantitatively determined by incubating the sample with solid-phase fixed antibodies against elastase first and reacting then with alkaline phosphatase-labelled antibodies against alpha 1-proteinase inhibitor. In normal plasma a level of 97.5 +/- 25.8 micrograms elastase/1 (mean +/- s.d., n = 43) was found, whereas moderately to markedly increased plasma concentrations were demonstrated in a variety of patients with inflammatory diseases like septicemia or rheumatoid arthritis.
Assuntos
Artrite Reumatoide/enzimologia , Proteínas Sanguíneas/metabolismo , Neutrófilos/enzimologia , Elastase Pancreática/sangue , Inibidores de Proteases/metabolismo , Sepse/enzimologia , Idoso , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Inflamação , Masculino , Pessoa de Meia-Idade , Elastase Pancreática/isolamento & purificação , Ligação Proteica , Valores de Referência , alfa 1-AntitripsinaRESUMO
The preparation of the aminomethyl-bilinogen which results from formal "head to tail" condensation of porphobilinogen is described. The chemical cyclocondensation of this compound at pH 7.4 yields uroporphyrinogen I. Enzymatic studies with enzyme preparations from Propionibacterium shermanii, which synthesize uroporphyrinogens from porphobilinogen, show that the rate of cyclisation is increased by these enzymes and indicate that the bilinogen also might be used for uroporphyrinogen III formation. This is also suggested by studies on the formation of cobyrinic acid from [4-14C]5-aminolevulinate via uroporphyrinogen III in the presence of the aminomethylbilinogen by cell-free extracts from Clostridium tetanomorphum.
