Post-translational Modifications (PTMs) Analysis

ICH Guideline Q6B states that protein/peptide biopharmaceuticals must be analyzed for post-translational modifications (PTMs) to characterize their purity and demonstrate batch consistency. With hundreds of successful projects in protein/peptide PTM analysis, CD Formulation provides unparalleled PTM analysis that performs as part of in-depth structural characterization studies and as an important component of comparability programs, stability studies, or quality control testing.

PTM & Why is PTM Analysis of Protein/peptide Important?

Post-translational modification (PTM) refers to the covalent modification of proteins, usually enzymatic modification, including phosphorylation, glycosylation, ubiquitination, nitrosylation, methylation, acetylation, lipidation, and proteolysis, which affects almost all aspects of normal cell biology and pathogenesis.

Protein/peptide PTMs is common in biopharmaceutical substances produced by bioprocessing. These PTM forms, also known as variants, have a certain degree of structural heterogeneity and are produced by adding functional groups (such as amide, phosphate, acetate, or methyl) after translation. PTM anal can affect the structure, stability, function, and interaction mode of proteins. Conducting PTM analysis can help to identify protein identity, ensure consistent manufacturing process and product quality, and identify erroneous modifications that may cause potential harmful immune responses in patients or reduce product efficacy, ensuring the safety and effectiveness of biopharmaceuticals.

Explore Our Post-translational Modifications (PTMs) Analysis Services

Determining protein/peptide post-translational modifications (e.g. phosphorylation, methionine oxidation, asparagine and glutamine deamidation, glycosylation changes, and proteolysis) requires a series of complex and detailed analyses to confirm protein/peptide identity and ensure consistent manufacturing processes and product quality.

As experts in the protein/peptide field and a full-service provider of PTM analysis services, CD Formulation's team of senior scientists can design strategic PTM analysis programs that meet regulatory requirements based on the specific characteristics of your product. Our scientists master the latest mass spectrometry analysis technology and methods to analyze various modifications such as phosphorylation, oxidation, asparagine and glutamine deamidation, glycosylation, and proteolysis.

We are able to identify and quantify a variety of possible post-translational modifications, including:

Fig.1 Typical PTMs of proteins/peptides. (CD Formulation)

• Phosphorylation: Ser/Thr, Tyr.
• Deamidation: Asn, Gln, Arg.
• Ubiquitination and SUMOylation: Lys, Gln.
• Oxidation: Histidine, tryptophan, and methionine.
• Methylation: Glu, Lys.
• Acylation: Acetylation, succinylation, myristoylation, palmitoylation, malonylation.
• Amino acid mutation: Selenocysteine to dehydroalanine, etc.
• Ribosylation: Gln, Asp, Lys.
• Glycosylation: Hydroxyproline.
• Nitration: Tyrosine.
• Non-natural amino acid chemical modifications.
• N/C-terminal sequenceing.

Our Solutions for Post-translational Modifications (PTMs) Analysis Services

Our biopharmaceutical analytical laboratories are equipped with state-of-the-art equipment and a broad base of expertise in providing you with high-resolution\accurate mass (HRAM) data on PTMs through mass spectrometry (MS).

In addition, our scientists use a range of chromatographic techniques to qualitatively identify and detect potential PTMs, and when necessary, use them in conjunction with mass spectrometry. Our capabilities cover:

Fig.2 Schematic illustration of bottom-up, middle-down, and top-down approaches for the identification of post-translational modifications.(Hermann J, et al., 2022)

PTM Identification

We offer comprehensive analysis services to identify and characterize various PTMs, including phosphorylation, acetylation, glycosylation, methylation, ubiquitination, and more, using state-of-the-art mass spectrometry techniques.

Site-specific PTM Localization

Our experts can accurately pinpoint the exact sites of PTMs on specific proteins, allowing for a deeper understanding of their functional roles and regulatory mechanisms.

Quantitative PTM Analysis

We provide quantitative analysis of PTMs to determine their abundance and dynamics in different experimental conditions or biological contexts, helping to unravel their roles in signaling pathways and disease mechanisms.

Bioinformatics Analysis

Our bioinformatics team can assist in the interpretation of PTM data, including pathway analysis, functional enrichment analysis, and integration with other omics data sets for a holistic view of PTM regulation.

PTM Validation

We offer validation services using orthogonal techniques such as western blotting, immunoprecipitation, and targeted mass spectrometry to validate and confirm the presence of specific PTMs.

Customized PTM Analysis

We can tailor our services to meet specific research needs, such as analyzing PTMs in specific protein targets or studying the effects of PTM-modifying enzymes or inhibitors.

Why Choose Us for PTM Analysis?

• We have extensive experience and expertise in performing various protein/peptide PTM analysis.
• Our PTM analysis services support multiple regulations, including USP, EP, JP, and PTC.
• We are able to accurately identify and quantify any PTMs, including phosphorylation, deamidation, glycosylation, ribosylation, etc.
• Our analytical laboratory is equipped with a range of state-of-the-art mass spectrometry equipment.
• We provide flexible and tailored solutions to meet the specific needs and requirements of our customers.
• Fast and efficient service ensures that customers' projects are delivered on time.

Publication

At CD Formulation, we understand the importance of protein and peptide PTMs analysis in the biologics development and manufacturing process. Please feel free to contact us if you are interested in our services. We will provide you with the most professional advice and support to ensure the smooth launch and implementation of your project.

How It Works

STEP 1

Submit a service request describing your specific research or development need.

STEP 2

We'll email you to provide your quote and confirm order details if applicable.

STEP 3

Execute the project with real-time communication, and deliver the final report promptly.