Identification of a Tetanus Toxin Specific Epitope in Single Amino Acid Resolution

Abstract

Vaccinations are among the most potent tools to fight infectious diseases. However, cross-reactions are an ongoing problem and there is an urgent need to fully understand the mechanisms of the immune response. For the development of a methodological workflow, the linear epitopes in the immune response to the tetanus toxin is investigated in sera of 19 vaccinated Europeans applying epitope mapping with peptide arrays. The most prominent epitope, appearing in nine different sera ( IHLVNNESSEVIVHK ), is investigated in a substitution analysis to identify the amino acids that are crucial for the binding of the corresponding antibody species - the antibody fingerprint. The antibody fingerprints of different individuals are compared and found to be strongly conserved ( ExxEVIVxK ), which is astonishing considering the randomness of their development. Additionally, the corresponding antibody species is isolated from one serum with batch chromatography using the amino acid sequence of the identified epitope and the tetanus specificity of the isolated antibody is verified by ELISA. Studying antibody fingerprints with peptide arrays should be transferable to any kind of humoral immune response toward protein antigens. Furthermore, antibody fingerprints have shown to be highly disease-specific and, therefore, can be employed as reliable biomarkers enabling the study of cross-reacting antigens.

Epitopes of Naturally Acquired and Vaccine‐Induced Anti‐Ebola Virus Glycoprotein Antibodies in Single Amino Acid Resolution

“…[ 31 ] Additionally, epitopes recognized by antibodies elicited after vaccination against infectious diseases, such as Tuberculosis, Malaria, and Tetanus, can be identified. [ 32–34 ]…”

A Low‐Cost Laser‐Based Nano‐3D Polymer Printer for Rapid Surface Patterning and Chemical Synthesis of Peptide and Glycan Microarrays

“…We show that it is possible to produce 9‐mer peptides with this simple setup, which is comparable to the high‐resolution LIFT method, reproducibly enabling a resolution of 250 µm (1600 spots cm −2 ). Peptide arrays are becoming increasingly important in research, and several applications in immunology and diagnostics have been shown recently, for example, the development of therapeutic antibodies or diagnostic biomarkers …”

Multivalent glycan arrays

“…Especially for peptide synthesis and applications in disease research, this offers a rapid strategy to generate diverse microarrays. [192][193][194][195][196][197] In the future, this technology may be exploited for the in situ synthesis of glycopeptides, glycans, and DNA in a microarray format.…”