Engineering β-lactamase zymogens for use in protease activity assays

2014 ◽  
Vol 50 (70) ◽  
pp. 10155-10157 ◽  
Author(s):  
Hajin Kim ◽  
Hyun Kyung Yoon ◽  
Tae Hyeon Yoo
Keyword(s):  
Protease Activity ◽  
Assay Method ◽  
Protease Assay

In this report, we engineered β-lactamase zymogens and developed a sensitive protease assay method based on the precursor enzymes.

A colorimetric protease activity assay method using engineered procaspase-3 enzymes

2016 ◽  
Vol 8 (33) ◽  
pp. 6270-6276 ◽  
Author(s):  
Dokyung Yang ◽  
Hyeon Ji Park ◽  
Tae Hyeon Yoo
Keyword(s):  
Protease Activity ◽  
Assay Method ◽  
Activity Assay ◽  
Platform Technology ◽  
Protease Assay ◽  
Diagnosis Of Diseases

A protease assay platform technology based on engineered proenzymes has been developed for diagnosis of diseases.

General Protease Assay Method Coupling Solid-Phase Substrate Extraction and Capillary Electrophoresis

1998 ◽  
Vol 70 (18) ◽  
pp. 3824-3827 ◽  
Author(s):  
Douglas B. Craig ◽  
Jerome C. Y. Wong ◽  
Robert Polakowski ◽  
Norman J. Dovichi
Keyword(s):  
Capillary Electrophoresis ◽  
Solid Phase ◽  
Assay Method ◽  
Protease Assay

Galactosidase Enzyme Fragment Complementation as a High-Throughput Screening Protease Technology

2004 ◽  
Vol 9 (5) ◽  
pp. 398-408 ◽  
Author(s):  
Tabassum Naqvi ◽  
Anice Lim ◽  
Riaz Rouhani ◽  
Raj Singh ◽  
Richard M. Eglen
Keyword(s):  
High Throughput ◽  
Protease Activity ◽  
High Throughput Screening ◽  
Caspase 3 ◽  
Cyclic Peptide ◽  
High Sensitivity ◽  
Enzyme Concentration ◽  
Protease Cleavage ◽  
Highly Sensitive ◽  
Protease Assay

The authors describe a homogeneous, high-throughput screening (HTS) assay for measuring protease activity and detection of inhibitors. The assay comprises a cyclic β-galactosidase (β-gal) enzyme donor peptide (ED) containing a protease-selective cleavage sequence. Alone, the cyclic peptide is inactive, but when linearized following protease cleavage, ED complements with β-gal enzyme acceptor forming active β-gal enzyme. This then catalyzes the formation of either fluorescent or chemiluminescent products, with β-gal turnover providing a highly amplified signal, and thus an assay technology of high sensitivity. To demonstrate the utility of the technology, an EFC assay was developed to measure the activity of 2, caspase 3 and β-secretase. Using a cyclic ED containing the caspase 3 substrate sequence, DEVD, the EFC assay signal was linear with respect to caspase 3 concentration. The assay was very sensitive, being able to detect activity at low picogram amounts of caspase 3. For the β-secretase (BACE) EFC assay, a cyclic ED containing the Swedish mutant cleavage site of amyloid precursor protein (APP), SEVNLDAEFK, was used. In a similar fashion to the caspase 3 assay, the signal induced by BACE activity was linear with respect to enzyme concentration and was highly sensitive, being able to detect nanogram quantities of BACE. The assay was also more sensitive than a commercially available FRET-based assay of BACE activity. It is concluded that the EFC protease assay is a simple, flexible, and sensitive technology for HTS of proteases.

Azocoll protease activity assay

2007 ◽  
Author(s):  
Naxin Jiang ◽  
Nguan Soon Tan ◽  
Bow Ho ◽  
Jeak Ling Ding
Keyword(s):  
Protease Activity ◽  
Activity Assay

The Measurement of Heparin

1973 ◽  
Vol 30 (03) ◽  
pp. 471-479 ◽  
Author(s):  
K. W. E Denson ◽  
John Bonnar
Keyword(s):  
Degradation Products ◽  
Factor Xa ◽  
Assay Method ◽  
Thrombin Time ◽  
Fibrinogen Degradation Products ◽  
Low Levels

SummaryA method for the measurement of heparin utilising the potentiating effect of heparin on the action of anti-factor Xa is described. The effect on the assay of platelet contamination of plasma, the presence of fibrinogen degradation products and low levels of anti-factor Xa have been studied. The assay method has been compared with the calcium thrombin time method and a group of obstetrical patients have been studied using both methods.

Sign in / Sign up

Export Citation Format

Share Document