Investigating Antibodies with Peptide Microarrays – Case Study
At axxelera, we are committed to supporting innovative research in the field of antibody screening.
Recently, our high-density Peptidome Arrays were utilized in a paper published in the International Journal of Molecular Sciences, focusing on selective binders to the therapeutic antibody rituximab.
Methodology
The objective of the study was to create a novel resemblance-ranking peptide library designed to closely mimic the human peptidome: the ensemble of all human proteins in the form of peptides.
High-density peptide microarrays displaying this library were used for a detailed multi-step screening process:
First, the researchers screened the peptide library to identify sequences that bind selectively to rituximab. Subsequently, they performed a substitutional analysis on the top 30 peptides to determine dissociation constants (KD) and identify invariant amino acids critical for binding.
This process enabled the identification of high-affinity peptide binders to the therapeutic antibody rituximab.
Key Findings
- Hydrophobic Amino Acids: The study found that the binding affinity of rituximab increased with the number of hydrophobic amino acids, particularly tryptophan and phenylalanine.
- Charge and Structure: Effective binders typically had low total charges and lower helix propensities.
- Dissociation Constants: The strongest binders had dissociation constants in the nanomolar range, indicating strong binding affinities.
Research Impact
The findings from this study advance our understanding of antibody interactions and epitope mapping, providing insights that are valuable for therapeutic antibody development.
They also demonstrate how peptide microarrays can play a crucial role in these discoveries, with their ability for large-scale and high-resolution screening.
Are you interested in how high-density peptide microarrays can support your research? Contact us today to discuss your project and learn more about our Peptidome Arrays.
