Protease and Peptidase Profiling Services

CD Biosynsis provides expert Protease and Peptidase Profiling Services crucial for drug discovery, biological research, and enzyme engineering. Proteases are a vast family of enzymes that control fundamental biological processes, making them significant therapeutic targets across various disease areas. Our integrated platform offers precise characterization of protease activity, inhibitor potency, and substrate specificity. We employ highly sensitive assays, including FRET-based, colorimetric, and fluorescence detection, to deliver quantitative data (Km, kcat, Ki, IC50). From profiling a novel therapeutic protease inhibitor to mapping the precise cleavage site of a new enzyme variant, our services ensure the accuracy and reliability required for confident decision-making. Select from our extensive library of validated proteases to construct a custom profiling panel.

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Advanced Tools for Understanding Peptide Cleavage

Protease profiling is complex due to the vast diversity in mechanism, substrate preference, and biological function across different protease classes (serine, cysteine, aspartic, metallo). Accurate characterization is necessary to distinguish specific therapeutic effects from unwanted off-target activities, a common challenge in drug development. Our service is built on a foundation of validated protease assays, high-throughput automation, and deep expertise in kinetic analysis. We are equipped to handle diverse targets, from pharmaceutical targets like Caspases and Cathepsins, to industrial enzymes. By providing flexible, customized target selection, we help you validate inhibitor mechanisms, optimize enzyme specificity, and ensure the stability of therapeutic protein drugs against degradation.

Protease Profiling Service Flow

A structured approach to comprehensive enzyme characterization.

Target Panel Selection and Sourcing

Assay Optimization and Validation

Kinetic and Inhibition Studies

Data Analysis and Mechanistic Reporting

Panel Customization: Finalize the list of Protease and Peptidase targets based on client selection and therapeutic focus.

Target Enzyme QC: Confirm purity, concentration, and baseline activity of the purified protease for all selected targets.

Assay Selection: Implement the most sensitive detection method (e.g., FRET, fluorescence polarization) for each target's substrate.

Optimization: Adjust pH, buffer composition, and enzyme concentration to achieve linear reaction kinetics and validate assay quality (Z-factor > 0.7).

IC50 Profiling: Run concentration-response curves for inhibitors for all selected targets to determine inhibitory strength (IC50).

Km/kcat Measurement: Perform full Michaelis-Menten kinetic runs for selected targets to define catalytic efficiency.

Mechanistic Analysis: Perform full kinetic studies to determine the inhibition constant (Ki) and inhibition mode.
Cleavage Site Confirmation: Use LC-MS/MS if requested to verify the precise site of peptide bond hydrolysis.
Reporting: Deliver all raw data, analysis plots, kinetic constants, and detailed technical methodology.

Precision and Specificity in Protease Analysis

Customizable Enzyme Panel

Freedom to select and combine targets from all major protease classes for highly specific profiling.

Substrate Specificity Mapping

Expert use of mass spectrometry and peptide libraries for definitive cleavage site identification and motif mapping.

Detailed Inhibition Kinetics

Accurate Ki determination and mechanistic distinction between reversible and time-dependent irreversible inhibitors.

High-Sensitivity Assays

Use of sensitive FRET and fluorescence probes for profiling low-activity or precious enzyme samples.

Client Testimonials on Protease Profiling Services

"The cleavage site mapping using mass spectrometry was critical. It definitively showed the off-target activity of our compound, allowing us to quickly redesign our peptide inhibitor."

Dr. Emily Zhao, Peptide Drug Discovery

"CD Biosynsis successfully adapted a challenging FRET assay for our new Cathepsin variant, delivering highly reproducible Km and kcat data essential for our patent application."

Mr. David Kim, Enzyme Engineering Lead

"The detailed Ki studies on our irreversible inhibitor provided the kinetic parameters (kinact/KI) needed for our preclinical studies, far exceeding the quality of our in-house data."

Ms. Sarah O'Connell, Pharmaceutical R&D

FAQs about Protease and Peptidase Profiling

Still have questions?

What is the P1' residue in protease substrate notation?

In protease substrate notation, the P1-P1' bond is the one cleaved. P1 is the residue on the N-terminal side of the cleavage site, and P1' is the first residue on the C-terminal side of the cleavage site.

How do you measure activity for metalloproteases?

Metallo-protease activity is commonly measured using FRET or colorimetric substrates, with careful control of the metal ion (typically Zn2+ or Ca2+) concentration, and using a chelator to distinguish non-specific activity.

Can you perform profiling on highly insoluble or membrane-associated proteases?

Yes, we have specialized protocols using detergents, liposomes, or lipid nanodiscs to maintain the activity and stability of challenging, membrane-associated, or poorly soluble proteases during the profiling assay.

What is the difference between IC50 and Ki in inhibition studies?

IC50 is the concentration of inhibitor that causes 50 percent inhibition of activity under specific assay conditions. Ki is the true equilibrium dissociation constant for the inhibitor-enzyme complex, determined through rigorous kinetic studies and independent of substrate concentration.

How much does Metabolic Engineering services cost?

The cost of Metabolic Engineering services depends on the project scope, complexity of the target compound, the host organism chosen, and the required yield optimization. We provide customized quotes after a detailed discussion of your specific research objectives.

Do your engineered strains meet regulatory standards?

We adhere to high quality control standards in all strain construction and optimization processes. While we do not handle final regulatory approval, our detailed documentation and compliance with best laboratory practices ensure your engineered strains are prepared for necessary regulatory filings (e.g., GRAS, FDA).

What to look for when selecting the best gene editing service?

We provide various gene editing services such as CRISPR-sgRNA library generation, stable transformation cell line generation, gene knockout cell line generation, and gene point mutation cell line generation. Users are free to select the type of service that suits their research.

Does gene editing allow customisability?

Yes, we offer very customised gene editing solutions such as AAV vector capsid directed evolution, mRNA vector gene delivery, library creation, promoter evolution and screening, etc.

What is the process for keeping data private and confidential?

We adhere to the data privacy policy completely, and all customer data and experimental data are kept confidential.

Protease and Peptidase Profiling Services

Advanced Tools for Understanding Peptide Cleavage

Protease and Peptidase Target Profiling Options

Selectable Serine and Cysteine Proteases

Choose Key Targets for Profiling

Selectable Metallo- and Aspartic Proteases

Choose Non-Serine/Cysteine Targets

Available Profiling Methods for Selected Targets

Assay Types and Kinetic Services

Protease Profiling Service Flow

Precision and Specificity in Protease Analysis

Client Testimonials on Protease Profiling Services

FAQs about Protease and Peptidase Profiling