Protein/Peptide Crosslinking

Chemical Crosslinking Services

Photoreactive Crosslinking Services

Enzymatic Crosslinking Services

With a comprehensive state-of-the-art platform, BOC Sciences is fully capable and committed to providing one-stop molecular bioconjugation services. BOC Sciences offers customers an integrated protein peptide binding service with crosslinking optimization and functional validation.

What is Protein Crosslinking?

Protein/peptide crosslinking refers to the formation of covalent bonds between proteins or peptides, resulting in the formation of complexes or network structures. This process can occur naturally in biological systems or can be artificially induced in the laboratory for different purposes. The structure of functional proteins involves more than just intramolecular covalent bonding, which leads to challenging questions about the spatial orientation and non-covalent interactions of proteins.

Fig.1 Online cross-linking of proteins. (Burris et al., 2022)

Protein Peptide Binding Services

BOC Sciences, a leading service provider in the field of drug discovery and research, can provide protein peptide binding services to clients worldwide to facilitate your research. We can perform protein or peptide crosslinking using a variety of methods, including chemical crosslinkers, photocrosslinking, and enzyme-catalyzed crosslinking.

(1) Chemical cross-linking

Reacting with amino groups (e.g., lysine residues) or carboxyl groups (e.g., aspartic acid or glutamic acid residues) via chemical cross-linking agents forms stable cross-links.

(2) Photocrosslinking

We also offer a photocrosslinking method for protein/peptide crosslinking. Photocrosslinking utilizes photosensitive molecules that are activated by light at specific wavelengths to produce reaction intermediates capable of forming covalent bonds with nearby proteins or peptides.

(3) Enzyme-catalyzed cross-linking

Enzyme-catalyzed cross-linking utilizes specific enzymes to catalyze the formation of cross-links between proteins or peptides. This method is used to generate protein-based materials or to change the structure of proteins.

• Services Available
  • Protein crosslinking
  • Peptide crosslinking
  • Protein peptide synthesis

Optimization of Protein Peptide Binding

• Selection of crosslinking reagents: Select the appropriate cross-linking reagent based on the desired cross-linking results and experimental goals.
• Concentration of cross-linking reagent: Different concentrations of cross-linking reagents can be tried, usually starting at lower concentrations and gradually increasing.
• pH value: Different proteins and crosslinking reagents may be pH sensitive. When optimizing experimental conditions, try different pH ranges to determine the best cross-linking effect.

Validation of Protein Peptide Binding

Determining the effect of a cross-linking reaction can be assessed by a variety of methods.

• Gel electrophoresis analysis: Gel electrophoresis can be used to assess the effectiveness of a cross-linking reaction. In gel electrophoresis, cross-linked proteins or peptides typically show higher molecular weights and form new bands or bands.
• Mass spectrometry: Mass spectrometry allows the identification and characterization of cross-linking products and the location of cross-linking sites.
• Western blotting: By immunoblotting cross-linked proteins or peptides, the presence of cross-linking products can be detected and their relative abundance determined.
• Functional evaluation: Depending on the purpose of the cross-linking, a functional assessment can be performed to determine the effectiveness of the cross-linking reaction. For example, if the cross-linking is intended to increase the stability of a protein or change its structure, the effect of the cross-linking can be assessed by assays of biological activity, stability, or structural properties.

Our Advantages

• Comprehensive and proficient bioconjugation technical platform
• All-round cross-linking chemistry
• Multiple methods for protein peptide binding validation
• Crosslinking analysis, detailed report with results and discussion

Project Workflow

Frequently Asked Questions (FAQ)

What methods are used for protein/peptide crosslinking?

Protein/peptide crosslinking can be performed using various techniques, including chemical crosslinking, photocrosslinking, and enzyme-catalyzed crosslinking. These methods allow for the creation of stable covalent bonds between proteins or peptides, each offering unique advantages depending on the desired application and experimental conditions.

How do you optimize the crosslinking process for proteins and peptides?

Optimization of protein/peptide crosslinking involves selecting the appropriate crosslinking reagent, adjusting the concentration of reagents, and fine-tuning the pH to achieve the best results. The process is carefully controlled to enhance binding efficiency while maintaining protein stability and functionality.

How can the effectiveness of crosslinking be validated?

The effectiveness of protein/peptide crosslinking can be validated through various techniques, including gel electrophoresis, mass spectrometry, and Western blotting. These methods help determine the molecular weight of the conjugates, identify crosslinking sites, and assess the overall success of the crosslinking reaction.