Expression, purification, biochemical and pharmacological characterization of a recombinant aprotinin variant

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dc.contributor.authorApeler H.
dc.contributor.authorPeters J.
dc.contributor.authorSchroder W.
dc.contributor.authorSchneider K.-H.
dc.contributor.authorLemm G.
dc.contributor.authorHinz V.
dc.contributor.authorRossouw G.J.
dc.contributor.authorDembowsky K.
dc.date.accessioned2011-05-15T16:15:39Z
dc.date.available2011-05-15T16:15:39Z
dc.date.issued2004
dc.description.abstractAprotinin (CAS 9087-70-1) is known as a potent inhibitor of serine proteases such as trypsin, plasmin, tissue and plasma kallikrein. In this study, an aprotinin variant was designed by means of rationale mutagenesis that differs from aprotinin by two amino acids in the active site and by seven amino acids in the backbone. The recombinant protein is expressed in a secretory yeast system enabling large scale production. A purification procedure was developed to yield high amounts of pure and correctly processed aprotinin variant. The changes in the active site of the aprotinin variant increase the potency towards inhibition of plasma kallikrein whereas the inhibition of plasmin is only marginally reduced. The net charge of the molecule is reduced from the basic (IP 10.5) to the neutral range (IP 5.6). The recombinant aprotinin variant shows a decrease of immunogenicity in several models. No cross-reactivity with human and rabbit antibodies directed against aprotinin was observed both in in vivo and in ex vivo studies. In addition, the variant is more potent in a rat brain edema model of acute subdural hematoma compared to aprotinin.
dc.description.versionArticle
dc.identifier.citationArzneimittel-Forschung/Drug Research
dc.identifier.citation54
dc.identifier.citation8
dc.identifier.issn00044172
dc.identifier.urihttp://hdl.handle.net/10019.1/13430
dc.subjectantiinflammatory agent
dc.subjectaprotinin
dc.subjectaprotinin variant
dc.subjectkallikrein
dc.subjectplasmin
dc.subjectrecombinant protein
dc.subjectserine proteinase inhibitor
dc.subjectunclassified drug
dc.subjectamino acid sequence
dc.subjectanimal experiment
dc.subjectanimal model
dc.subjectanimal tissue
dc.subjectarticle
dc.subjectbrain edema
dc.subjectbrain water
dc.subjectchimpanzee
dc.subjectcontrolled study
dc.subjectcross reaction
dc.subjectdog
dc.subjectdrug potency
dc.subjectdrug purification
dc.subjectenzyme active site
dc.subjectenzyme inhibition
dc.subjectex vivo study
dc.subjectfemale
dc.subjecthuman
dc.subjecthuman cell
dc.subjectimmunogenicity
dc.subjectin vitro study
dc.subjectin vivo study
dc.subjectinhibition kinetics
dc.subjectmale
dc.subjectnonhuman
dc.subjectprotein expression
dc.subjectrabbit
dc.subjectrat
dc.subjectreflex
dc.subjectSaccharomyces cerevisiae
dc.subjectsite directed mutagenesis
dc.subjectsubdural hematoma
dc.subjectsurface charge
dc.subjectAmino Acids
dc.subjectAnimals
dc.subjectAprotinin
dc.subjectBody Water
dc.subjectBrain Chemistry
dc.subjectBrain Edema
dc.subjectChemistry, Physical
dc.subjectChromatography, High Pressure Liquid
dc.subjectCloning, Molecular
dc.subjectCross Reactions
dc.subjectDNA, Complementary
dc.subjectDogs
dc.subjectElectrophoresis, Capillary
dc.subjectElectrophoresis, Polyacrylamide Gel
dc.subjectFemale
dc.subjectFermentation
dc.subjectFreeze Drying
dc.subjectHand Strength
dc.subjectHemodynamic Processes
dc.subjectHistamine Release
dc.subjectIsoelectric Focusing
dc.subjectMale
dc.subjectMolecular Weight
dc.subjectPan troglodytes
dc.subjectPeptide Mapping
dc.subjectProtease Inhibitors
dc.subjectRats
dc.subjectRats, Wistar
dc.subjectRecombinant Proteins
dc.subjectSaccharomyces cerevisiae
dc.subjectSequence Analysis, Protein
dc.titleExpression, purification, biochemical and pharmacological characterization of a recombinant aprotinin variant
dc.typeArticle
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