“PMN-Elastase Assay”: Enzyme Immunoassay for Human Polymorphonuclear Elastase Complexed with α1-Proteinase Inhibitor

Summary:We investigated the diagnostic significance of UDP-Z>-xylose: proteoglycan core protein ß-£>-xylosyltransferase (EC 2.4.2.26) in different chronic joint diseases. This eiizyme is located almost exclusively within chondrocytes, where it initiales the formation of chondroitin sulphate during the biosynthesis of proteoglycans and from which it is easily released after damage of articular cartilage.

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“…Elastase in complex with a r proteinase inhibitor was determined by an enzyme immunoassay described previously (18).…”

Section: Other Methodsmentioning

confidence: 99%

UDP-D-Xylose : Proteoglycan Core Protein β-D-Xylosyltransferase: A New Marker of Cartilage Destruction in Chronic Joint Diseases

Kleesiek

Reinards²,

Okusi³

et al. 1987

Clinical Chemistry and Laboratory Medicine

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“…In vivo extracellularly discharged PMN-E is rapidly complexed by the plasma inhibitors el-proteinase inhibitor (90%) and c~2-macroglobulin (10%). Therefore, in biological fluids, especially in plasma, elastase is measured primarily as the inactive enzyme in complex with cq-proteinase inhibitor [12,20]. Yet, in inflammatory local body fluids the inhibitory potential may be consumed to such a degree that proteolytically active elastase is also present [1,13].…”

Section: Discussionmentioning

confidence: 99%

“…PMN-E in synovial fluids in complex with cdproteinase inhibitor (cdPI) was determined according to Neumann [20] using a specific two-site sandwich ELISA (Merck, Darmstadt, Germany). Moreover, to detect proteolytically active elastase which may not be inhibited in the local inflammatory environment due to lack of sufficient inhibitory active ~aPI, one part of each synovial fluid was mixed with a surplus of purified c~lPI and reassayed for an in vitro increase of the elastase-elPI complex.…”

Section: Methodsmentioning

confidence: 99%

Differential patterns of PMN-elastase and type III procollagen peptide in knee joint effusions due to acute and chronic sports injuries

et al. 1991

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Summary.In 38 traumatic knee joint effusions the proteolytic enzyme PMN-elastase (PMN-E) and the repair marker procollagen III aminoterminal peptide (PIIINP) were determined. According to the period between trauma and first aspiration of the effusion, the patients were divided into 3 groups. Group I (17 patients; period between trauma and first aspiration not longer than 72 hours) showed high concentrations of PMN-E (up to 5400 ng/ml) and low concentrations of PIIINP (< 13 U/ml). Group II (11 patients; aspiration within 4 to 14 days) had mean PMN-E and PIIINP concentrations of 125.6 ng/ml and 52.1 U/ ml, respectively. In group III (10 patients, aspiration after 14 days) mean PMN-E concentration was 123.8 ng/ml and mean PIIINP concentration was 63.4 U/ml. Graphic depiction of PMN-E and PIIINP levels in each individual sample as a function of time between trauma and fluid collection revealed highly increasing PMN-E levels during the first 24 posttraumatic hours, followed by rapidly decreasing levels within 72 hours post trauma, and no change after the 4th posttraumatic day. In contrast, PIIINP increased continuously up to the first posttraumatic week and stayed 'at high levels up to 90 days (end of the observation period). The differential patterns of PMN-E and PIIINP concentration in knee joint effusions may be useful in estimating the period between trauma and first treatment (aspiration of effusion) and should, therefore, be helpful in detecting degenerative lesions, which seem to be characterized by low PMN-E concomitantly with high PIIINP levels.

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“…For the enzyme immunoassay a p rocedure as repo rted p reo~s!y fo r tbe compJell of PMN elastase Wilh Cl.1 -proteinase mh!bltor was followed analogously [6]. Brieny, polystyrcne tubcs (10.5 mm x 40 mm) werc coated by incubation wilh O.5 ml containing 50 ).lg sheep T gG 10 MPOJml in p hosphate buffered saline pH 7.5 (PBS) at room te mperature (20 to 2rq ovem ight and washed with 0.05% (wJv) polyoxyethylene (20)-sorbitan monolaurate scveral ti mes before use.…”

Section: Materials and Mcthodsmentioning

confidence: 99%

“…T he IgG fractions were isolated by ammonium sulfate precipitation and ion-exchange chromatography on O EAE cellulose. The specific IgG fractioD from rabbilS was isolatcd by adding immunoadsorption on MPO-agarose, and thcn coupled 10 calf intestinal al kaline pbosphatase with glutardialdehyde as describcd previously {5].For the enzyme immunoassay a p rocedure as repo rted p reo~s!y fo r tbe compJell of PMN elastase Wilh Cl.1 -proteinase mh!bltor was followed analogously [6]. Brieny, polystyrcne tubcs (10.5 mm x 40 mm) werc coated by incubation wilh O.5 ml containing 50 ).lg sheep T gG 10 MPOJml in p hosphate buffered saline pH 7.5 (PBS) at room te mperature (20 to 2rq ovem ight and washed with 0.05% (wJv) polyoxyethylene (20)-sorbitan monolaurate scveral ti mes before use.…”

mentioning

confidence: 99%

Quantitation of myeloperoxidase from human granulocytes as an inflammation marker by enzyme-linked immunosorbent assay

et al. 1986

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Federal Rep ublic of Germany Q uantifizierung von Myeloperoxidase aus humanen Granulocyten als [ntlündungsparameter mittels J::nzymimmunoassay \'om SandwiehlYP Myeloperoxidase (donor: H 2 0 2 oxidorcd uctase, EC 1.11.1. 7) is a cationic heme enzyme of mol wt of approximately 140,000 whieh is found as a main constituent of the azurophilic granules of polymorphonuclcar lcukoeytes (PMN) and in monocytcs (1]. Myclopcroxidase (MPO) plays an important role in killing bacteria using halide ions as a cofactor [2]. As Materials and mcthodsMPQ was purified from nOnTIal human granulocytes by a procedurc involving acid elltraction of gran ules, copper chelate chromat0f::aphy, cation exchange chromatography on SPSephadex M C-50 and size exclusion chromatography on Fractogelll. TId TSK HW·55 . Tbc final enzyme prepamtion was sho:wn to bc ffee of oth~r granular proteins by SOS-polyacrylamIde gel electrophores!s and immunoelectrophoresis against various polyvalent or monospecifi c autisera against granular components. -Antigen.spccific anlisera to MPO were raised in sheep and rabbilS. T he IgG fractions were isolated by ammonium sulfate precipitation and ion-exchange chromatography on O EAE cellulose. The specific IgG fractioD from rabbilS was isolatcd by adding immunoadsorption on MPO-agarose, and thcn coupled 10 calf intestinal al kaline pbosphatase with glutardialdehyde as describcd previously {5].For the enzyme immunoassay a p rocedure as repo rted p reo~s!y fo r tbe compJell of PMN elastase Wilh Cl.1 -proteinase mh!bltor was followed analogously [6]. Brieny, polystyrcne tubcs (10.5 mm x 40 mm) werc coated by incubation wilh O.5 ml containing 50 ).lg sheep T gG 10 MPOJml in p hosphate buffered saline pH 7.5 (PBS) at room te mperature (20 to 2rq ovem ight and washed with 0.05% (wJv) polyoxyethylene (20)-sorbitan monolaurate scveral ti mes before use. FO T tbe assay 0.5 ml sampie or calibrator was incubated in the coated tubes for I h at 20 10 22°C. Tbe tubes were washed three times, and 0.5 ml enzyme labellcd anlibodies to MPO in saJine Tris/HCI buffer pH 8.0 was added . After incubation for 2 h at 20 to 22~C and wa~h i ng of the tubes 0.5 ml substrate soLution, i.c. 10 mmol/l 4-nitrophenyl phosphate in I molfl diethanolamine/HCI buffer pH 9.8 witb 0.5 mmol MgCI 2 /1, was dclivered to each tube and incubated for 1 hat 20 to 2rc. 0.5 ml 2 mol/l NaOH wasaddcd witb vortexing. Absorbance was read at 405 nm . A40s was plotted agains t MPO concentration on a linear seale. As a rou tine citrated pL asma was used in a final dilution of one part plasma plus 20 or 50 parts of PBS containing 1 mg bovine serum albumin/mi and 20 mmol EOTA/I. Calibrators wcre used in tbe same diluent pL us 2"/" (viv) citrated normal sheep plasma_ Results and discussion A linear calibration curve was obtained for thc range of 0 to 20llg MPO/ I. Unk nowns werc detcrmined by reference 10 the calibration curve as calculated by linear regression analysis. Detection limit, i.e. mean absorbance of tbe blank plus 3 x standard deviation, was abOllI 0.25 ).Ig/l....

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“PMN-Elastase Assay”: Enzyme Immunoassay for Human Polymorphonuclear Elastase Complexed with α1-Proteinase Inhibitor

Cited by 91 publications

References 10 publications

UDP-D-Xylose : Proteoglycan Core Protein β-D-Xylosyltransferase: A New Marker of Cartilage Destruction in Chronic Joint Diseases

UDP-D-Xylose : Proteoglycan Core Protein β-D-Xylosyltransferase: A New Marker of Cartilage Destruction in Chronic Joint Diseases

Differential patterns of PMN-elastase and type III procollagen peptide in knee joint effusions due to acute and chronic sports injuries

Quantitation of myeloperoxidase from human granulocytes as an inflammation marker by enzyme-linked immunosorbent assay

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