Description:
Mapping targets of antibodies - in particular neutralizing ones - is of great value for monitoring vaccination efficacy as well as for development of novel vaccines and reagents for diagnostics. To this end, several methods (e.g. phage display libraries, peptide-based ELISA) have been used. These, however, suffer from cumbersome handling and the need of large amounts of sera, often pooled from many patients with the effect of blinding the serological analysis to individual variation. As shown here, these disadvantages can be overcome by miniaturization and parallelization. In this study, hepatitis B virus (HBV) and human immunodeficiency virus envelop (HIVenv) chips with overlapping oligopeptides encompassing the full amino acid sequences of different HBV and HIV polypeptides were produced via SPOT synthesis (Frank 1992) and printing to glass slides (Dikmans, Beutling et al. 2006). In addition, a chip displaying a library of 4608 different 15-mers of defined but randomly chosen sequence (4608-RPL) was prepared. HBV and HIVenv chips were used for analyzing monoclonal antibodies, serum samples from HBV vaccinated or infected individuals, and sera from HIV/SIV-immunized monkeys. Both chips allowed for high-resolution mapping of B cell linear epitopes. The identified epitopes were 3, 6, 9 or 12 amino acids in length. Moreover, it was shown that, for a great extent, the 4608-RPL could be used for identifying target sequences of monoclonal antibodies detecting several types of B cell epitope. Dikmans, A., U. Beutling, et al. (2006). "SC2: A novel process for manufacturing multipurpose high-density chemical microarrays." Qsar & Combinatorial Science 25(11): 1069-1080. Frank, R. (1992). "Spot-Synthesis - an Easy Technique for the Positionally Addressable, Parallel Chemical Synthesis on a Membrane Support." Tetrahedron 48(42): 9217-9232.