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Peparray™ Antibody Repertoire Analysis

As a long-term expert in applying phage display technology and peptide array technology, Creative Biolabs has accumulated extensive experiences. Combining this two advanced technologies, Creative Biolabs can provide high-quality service for single amino acid fingerprinting of antibody repertoire. With our strategy, it is possible to pinpoint those antibodies that correlate with a certain antigen, without any pre-information. We are proud to share our leading technology to facilitate the research and project development of all our customers.

Antibody species are selected from a plethora of different antibodies. Most of these selected antibody species are further refined in their binding characteristics by random point mutations and somatic hypermutation. There have been many different approaches to analyze the diversity of a patient’s individual antibody repertoire, such as high-throughput sequencing techniques. However, these techniques rely on the conclusion that the full antibody repertoire can be deduced from the gene expression at a certain time point. This expression might be somewhat different to the actual diversity and specificities of antibodies. In today's typical antibody-based diagnostics, the disease status is concluded from the binding of a patient’s serum antibodies to one or a few proteins. Immunoblotting techniques confirm that only a limited number of proteins of a pathogen are targeted. Thus, the immune systems of different individuals seem to commonly prefer only a few proteins out of thousands of potential antigens of a bacterial pathogen or in autoimmune disease. Whether the immune systems of different patients target the same proteins or similar epitopes, these basic scientific questions are still unanswered. Simultaneously, we would need to know exactly which amino acids are crucial for the binding of many different antibody species.

Fig.1 Validation by peptide array after prescreening by phage display (Weber et al. 2017).
Fig.1 Validation by peptide array after prescreening by phage display (Weber et al. 2017).

To resolve this question, Creative Biolabs combined our phage display platform with Peparray™ high throughput screening (HTS) peptide array platform for single amino acid fingerprinting of the antibody repertoire. In an initial pre-screen, up to 109 random peptides displayed on phage were screened for their binding to serum antibodies. Next, the identified epitope peptides were validated on our advanced Peparray™ HTS peptide array platform. Finally, the validated epitopes were fine mapped by comprehensive substitution analysis. The resulting “epitope fingerprints” enable the identification of those proteins that match the antibody.

Key features


Creative Biolabs has rich experience in single amino acid fingerprinting of the antibody repertoire for key residues identification. This approach allows us to evaluate the specificity of an antibody down to the individual amino acid level and refines the profiling of the antibody repertoire. Besides gaining knowledge on the antibody repertoire, our approach is useful for elucidating clinical phenotypes with unknown disease antigens, without any prior knowledge. We are confident in providing clients with the best service at the most competitive cost.

For more detailed information, please feel free to contact us or directly send us an inquiry.

Reference

  1. Weber, L.K.; et al. Single amino acid fingerprinting of the human antibody repertoire with high-density peptide arrays. J Immunol Methods. 2017, 443:45-54.



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