scholarly journals Sensitive and specific measurement of alpha1-antitrypsin activity with an elastase complex formation immunosorbent assay (ECFISA)

2021 ◽  
pp. 114476
Author(s):  
Andrea Engelmaier ◽  
Alfred Weber
Keyword(s):  
Complex Formation ◽  
Immunosorbent Assay ◽  
Specific Measurement ◽  
Alpha1 Antitrypsin ◽  
Antitrypsin Activity

Alpha1-antitrypsin activity in subarachnoid hemorrhage

Life Sciences ◽  
1996 ◽  
Vol 59 (1) ◽  
pp. 15-20 ◽  
Author(s):  
Fulvio Tartara ◽  
Paolo Gaetani ◽  
Flavio Tancioni ◽  
Alberto Guagliano ◽  
Catherine Klersy ◽  
...  
Keyword(s):  
Subarachnoid Hemorrhage ◽  
Alpha1 Antitrypsin ◽  
Antitrypsin Activity

Altered elastase—alpha1-antitrypsin balance in the blood of patients with chronic venous disease

2015 ◽  
Vol 31 (2) ◽  
pp. 125-132 ◽  
Author(s):  
M Budzyn-Napierala ◽  
M Iskra ◽  
Z Krasinski ◽  
W Turkiewicz ◽  
B Gryszczynska ◽  
...  
Keyword(s):  
Clinical Symptoms ◽  
Early Stage ◽  
Chronic Venous Disease ◽  
Venous Disease ◽  
Leukocyte Elastase ◽  
Vein Wall ◽  
Alpha1 Antitrypsin ◽  
Antitrypsin Activity ◽  
Wall Injury

Objectives Although leukocyte elastase is suspected to be involved in the damage of vein wall during chronic venous disease, the equilibrium between this protease and its inhibitor, alpha1-antitrypsin, has not yet been evaluated. The aim of the present study was to determine the relationship between leukocyte elastase and alpha1-antitrypsin, in the blood of patients with chronic venous disease. Patients and methods The concentration and the activity of leukocyte elastase along with the activity of alpha1-antitrypsin were evaluated in the blood of 55 chronic venous disease patients. The results were compared with those obtained in 33 healthy age and sex-matched volunteers. Results A significant decrease in the leukocyte elastase activity that correlated with an increased alpha1-antitrypsin activity was observed in the serum of patients with mild clinical symptoms of chronic venous disease. Conclusions The results of the study did not confirm a hypothesis about an important role of proteolytic activity of leukocyte elastase in the vein wall injury mechanism. They show that the leukocyte elastase–alpha1-antitrypsin balance is rather shifted toward antiprotease activity, especially in an early stage of chronic venous disease.

In vivo complex formation of modified alpha1-antitrypsin with LDL

2000 ◽  
Vol 151 (1) ◽  
pp. 54
Author(s):  
S Mashiba ◽  
Y Wada ◽  
M Takeya ◽  
A Sugiyama ◽  
T Hamakubo ◽  
...  
Keyword(s):  
Complex Formation ◽  
Alpha1 Antitrypsin

scholarly journals Assessment of Hageman factor activation in human plasma: quantification of activated Hageman factor-C1 inactivator complexes by an enzyme- linked differential antibody immunosorbent assay

Blood ◽  
1985 ◽  
Vol 66 (3) ◽  
pp. 636-641
Author(s):  
AP Kaplan ◽  
B Gruber ◽  
PC Harpel
Keyword(s):  
Complex Formation ◽  
Immunosorbent Assay ◽  
Time Dependent ◽  
Enzyme Linked Immunosorbent Assay ◽  
Contact Activation ◽  
Hageman Factor ◽  
Normal Plasma ◽  
Dose Dependent Increase ◽  
Dose Dependent

An enzyme-linked immunosorbent assay has been developed for the quantitation of activated Hageman factor-C1 inactivator (HF-C1 INH) complexes. Addition of increasing quantities of either of the major forms of activated Hageman factor (HFa or HFf) to normal plasma or to Hageman factor-deficient plasma leads to a dose-dependent increase in activated HF-C1 INH complexes. As little as 0.5 micrograms/mL of activated HF added to plasma can be detected, corresponding to activation of approximately 2% of plasma HF. The sensitivity of the assay is increased at least tenfold when complexes are formed in HF- deficient plasma, indicating competition between unactivated HF and activated HF-C1 INH complexes for binding to the antibody. Specificity is demonstrated in that addition of activated HF to hereditary angioedema plasma yields less than 1% of the activated HF-C1 INH complex formation obtained with normal plasma. Kaolin activation of HF- deficient plasma yields no detectable complex formation. Kaolin activation of prekallikrein-deficient plasma demonstrates a time- dependent increase in formation of activated HF-C1 INH complex consistent with the ability of HF in this plasma to autoactivate as the time of incubation with the surface is increased. Kaolin treatment of high-molecular weight (HMW) kininogen-deficient plasma yields an even more profound abnormality in the rate of formation of activated HF-C1 INH complexes reflecting the complex role of HMW kininogen in the initiation of contact activation. Although addition of corn inhibitor to plasma prevents activated HF-C1 INH complex formation, it does not inhibit activated HF sufficiently fast to prevent prekallikrein activation.

scholarly journals Identification of and partial characterization of platelet vitronectin: evidence for complex formation with platelet-derived plasminogen activator inhibitor-1

Blood ◽  
1989 ◽  
Vol 74 (6) ◽  
pp. 1989-1996 ◽  
Author(s):  
KT Preissner ◽  
S Holzhuter ◽  
C Justus ◽  
G Muller-Berghaus
Keyword(s):  
Complex Formation ◽  
Plasminogen Activator ◽  
Immunosorbent Assay ◽  
Plasminogen Activator Inhibitor ◽  
Enzyme Linked Immunosorbent Assay ◽  
Plasminogen Activator Inhibitor 1 ◽  
Platelet Factor ◽  
Adhesive Proteins ◽  
Activator Inhibitor ◽  
Pai 1

Abstract Vitronectin (VN; = complement S-protein), a plasma glycoprotein that is also associated with extracellular sites, was identified in washed human platelets contaminated with less than 0.05% of plasma VN. A specific enzyme-linked immunosorbent assay (ELISA) for VN has been developed and was used to detect and to quantitate VN in detergent extracts of washed platelets with 8.1 +/- 4.6 micrograms/10(9) platelets (n = 10), representing about 0.8% of the plasma VN pool. Platelet and plasma VN were similar by immunochemical criteria using Western-blot analysis, although platelet VN was mainly found as partially proteolyzed polypeptide. Total release of platelet VN occurred at optimal doses of Ca-ionophore 23187 or thrombin, whereas no VN was released by platelet treatment with digitonin or Staphylococcus alpha-toxin. During stimulation of washed platelets with various concentrations of thrombin, the nearly concomitant release of VN and plasminogen activator inhibitor-1 (PAI-1) together with platelet factor 4 indicated the association of VN with inner-platelet storage granules. Furthermore, platelet VN and PAI-1 in Ca-ionophore releasates comigrated during ultracentrifugation in high mol wt fractions of sucrose density gradients, indicating a possible association of both components. Complex formation of platelet VN and PAI-1 was verified by a sensitive enzyme-linked immunosorbent assay (ELISA) and accounts at least in part for a high molecular form of platelet VN. The identification of platelet VN and its binding to platelet PAI-1 raises the possibility that VN, in contrast to other adhesive proteins, may participate in localized regulatory functions of blood coagulation and fibrinolysis in platelet-matrix interactions and the protection of the matrix against proteolysis.

scholarly journals Assessment of Hageman factor activation in human plasma: quantification of activated Hageman factor-C1 inactivator complexes by an enzyme- linked differential antibody immunosorbent assay

Blood ◽  
1985 ◽  
Vol 66 (3) ◽  
pp. 636-641 ◽  
Author(s):  
AP Kaplan ◽  
B Gruber ◽  
PC Harpel
Keyword(s):  
Complex Formation ◽  
Immunosorbent Assay ◽  
Time Dependent ◽  
Enzyme Linked Immunosorbent Assay ◽  
Contact Activation ◽  
Hageman Factor ◽  
Normal Plasma ◽  
Dose Dependent Increase ◽  
Dose Dependent

Abstract An enzyme-linked immunosorbent assay has been developed for the quantitation of activated Hageman factor-C1 inactivator (HF-C1 INH) complexes. Addition of increasing quantities of either of the major forms of activated Hageman factor (HFa or HFf) to normal plasma or to Hageman factor-deficient plasma leads to a dose-dependent increase in activated HF-C1 INH complexes. As little as 0.5 micrograms/mL of activated HF added to plasma can be detected, corresponding to activation of approximately 2% of plasma HF. The sensitivity of the assay is increased at least tenfold when complexes are formed in HF- deficient plasma, indicating competition between unactivated HF and activated HF-C1 INH complexes for binding to the antibody. Specificity is demonstrated in that addition of activated HF to hereditary angioedema plasma yields less than 1% of the activated HF-C1 INH complex formation obtained with normal plasma. Kaolin activation of HF- deficient plasma yields no detectable complex formation. Kaolin activation of prekallikrein-deficient plasma demonstrates a time- dependent increase in formation of activated HF-C1 INH complex consistent with the ability of HF in this plasma to autoactivate as the time of incubation with the surface is increased. Kaolin treatment of high-molecular weight (HMW) kininogen-deficient plasma yields an even more profound abnormality in the rate of formation of activated HF-C1 INH complexes reflecting the complex role of HMW kininogen in the initiation of contact activation. Although addition of corn inhibitor to plasma prevents activated HF-C1 INH complex formation, it does not inhibit activated HF sufficiently fast to prevent prekallikrein activation.

scholarly journals Identification of and partial characterization of platelet vitronectin: evidence for complex formation with platelet-derived plasminogen activator inhibitor-1

Blood ◽  
1989 ◽  
Vol 74 (6) ◽  
pp. 1989-1996
Author(s):  
KT Preissner ◽  
S Holzhuter ◽  
C Justus ◽  
G Muller-Berghaus
Keyword(s):  
Complex Formation ◽  
Plasminogen Activator ◽  
Immunosorbent Assay ◽  
Plasminogen Activator Inhibitor ◽  
Enzyme Linked Immunosorbent Assay ◽  
Plasminogen Activator Inhibitor 1 ◽  
Platelet Factor ◽  
Adhesive Proteins ◽  
Activator Inhibitor ◽  
Pai 1

Vitronectin (VN; = complement S-protein), a plasma glycoprotein that is also associated with extracellular sites, was identified in washed human platelets contaminated with less than 0.05% of plasma VN. A specific enzyme-linked immunosorbent assay (ELISA) for VN has been developed and was used to detect and to quantitate VN in detergent extracts of washed platelets with 8.1 +/- 4.6 micrograms/10(9) platelets (n = 10), representing about 0.8% of the plasma VN pool. Platelet and plasma VN were similar by immunochemical criteria using Western-blot analysis, although platelet VN was mainly found as partially proteolyzed polypeptide. Total release of platelet VN occurred at optimal doses of Ca-ionophore 23187 or thrombin, whereas no VN was released by platelet treatment with digitonin or Staphylococcus alpha-toxin. During stimulation of washed platelets with various concentrations of thrombin, the nearly concomitant release of VN and plasminogen activator inhibitor-1 (PAI-1) together with platelet factor 4 indicated the association of VN with inner-platelet storage granules. Furthermore, platelet VN and PAI-1 in Ca-ionophore releasates comigrated during ultracentrifugation in high mol wt fractions of sucrose density gradients, indicating a possible association of both components. Complex formation of platelet VN and PAI-1 was verified by a sensitive enzyme-linked immunosorbent assay (ELISA) and accounts at least in part for a high molecular form of platelet VN. The identification of platelet VN and its binding to platelet PAI-1 raises the possibility that VN, in contrast to other adhesive proteins, may participate in localized regulatory functions of blood coagulation and fibrinolysis in platelet-matrix interactions and the protection of the matrix against proteolysis.

The Interaction of Proline-Rich Ligands with Profilin Probed with an Enzyme-Linked Immunosorbent Assay

2009 ◽  
Vol 14 (4) ◽  
pp. 350-359 ◽  
Author(s):  
Sylvie Veniere ◽  
Christophe Ampe ◽  
Joël Vandekerckhove ◽  
Anja Lambrechts
Keyword(s):  
Complex Formation ◽  
Ligand Binding ◽  
Immunosorbent Assay ◽  
Ternary Complex ◽  
Screening Method ◽  
Antibody Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Ternary Complex Formation

To detect interactions of different proline-rich ligands with profilins, the authors developed a simple analytical antibody-based screening method. Profilin I or profilin IIa was coated in microplates, and ligand binding was monitored via antibody detection. Using purified components, the authors show that the assay is very sensitive as nanomolar concentrations of recombinant profilin ligands can be used. They further apply this technique to detect interaction of profilin with various proline-rich partners, either endogenously present or ectopically expressed as tagged fusions, using lysates. With this assay, the authors identify Shootin1 as a novel profilin IIa partner. In addition, they demonstrate that this assay can be used for studying competition or ternary complex formation. In conclusion, they developed a sensitive, easy-to-use, and versatile method for the study of the interaction between profilin and different ligands. ( Journal of Biomolecular Screening 2009:350-359)

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