Steady-state enzyme kinetics is a cornerstone technique in biochemistry and related sciences because it allows the characterization and quantification of enzyme behavior. Enzyme kinetics is widely used to study the physiological effects of enzymes, to determine the effects of mutations, and to characterize enzyme inhibitors. Well-known examples of enzyme inhibitors used to treat diseases include anti-infectives; anti-inflammatory drugs (e.g., aspirin and ibuprofen); cholesterol-lowering statins; and tyrosine kinase inhibitors used to treat cancer. Often, new treatments for diseases can be discovered by using enzyme kinetics to identify the few active compounds present in large collections of compounds.Creative Enzymes services, including method development, validation, transfer and method development, help you accelerate your drug development process based on your every need!

Custom Cloning Enzyme Steady-State Kinetic Measurements Approaches We Offer

• Measurement of dose-response curves and IC 50 values

The first step in determining inhibitor activity is usually to determine the dose-response curve. Most inhibitors will bind to the enzyme in less than 1 second, but the binding of inhibitors can be much slower than this. Many inhibitors have limited solubility in the buffer and tend to precipitate, so a concentrated inhibitor stock solution should be prepared in DMSO and mixed with a larger volume of enzyme stock solution. At the end of the pre-incubation period, split the sample into replicates, add the substrate solution and measure the rate.

• Determine if the inhibitor is reversible

Proper characterization of an inhibitor requires determining whether it binds reversibly to the enzyme. We use rapid dilution experiments to determine this.

• Determination of K i values

The K i value is related to the thermodynamic strength of reversible inhibitor binding and is used to compare the potency between different reversible inhibitors. The most common method for determining K i values is to measure enzyme activity in the presence of variable substrate concentrations and several fixed inhibitor concentrations. We use several plots to analyze the data, including direct linear plots, direct ('Michaelis-Menten') plots, and various linear transformations of the Michaelis-Menten equation.

Service Process

Our Features

• Creative Enzymes has established a proven enzyme technology platform for a variety of applications. We'll help you design the right cloning enzyme steady-state kinetic measurements service to aid your scientific research.
• Creative Enzymes is a leading enzyme technology company offering a trusted suite of services and solutions, especially with our extensive experience in cloning enzyme steady-state kinetic measurements service.

Deliverables

• Results raw data
• Data analysis
• Results analysis
• Purification assessment report
• Summary of relevant parameters

Why Choose Us?

Enzyme inhibitors are the largest class of small molecule drugs used in modern human and veterinary medicine and have been very successful. Novel enzyme inhibitors are being actively developed for the treatment of many different diseases. For the foreseeable future, enzyme kinetics will remain a key technology for characterizing inhibitors, optimizing drug efficacy, and identifying inhibitors through high-throughput screening and related technologies.Creative Enzymes is a professional cloning enzyme steady-state kinetic measurements service provider supporting a wide range of enzyme services. We are able to perform individual tests at the most precise level, as well as design and complete a suite of services as a solution to the subject project.