Epitope Mapping of potential novel immunogenic proteins of Klebsiella pneumoniae MGH 78578. Klebsiella pneumoniae

A number of six proteins were selected during immunoscreening and further analyses. The proteins were divided in silico into overlapping 15-mer oligopeptides with an overlap of 11 residues. The microarrays were incubated with different antibodies to K. pneumoniae, C. jejuni and S. aureus. Overall design: Each microarray was seperated into three individual incubation chambers using ProPlate 3-Well modules. Within each incubation chamber each peptide was spotted in triplicate with the controls spotted nine times each. The controls included Human-IgG, Rabbit-IgG, Mouse-IgG and Myelin Basal Protein. Each chamber was incubated independently using different polyclonal antibodies to K. pneumoniae and for specificity testing with antibodies to C. jejuni or S. aureus. Thus, samples 2_1, 2_2, 3_1 and 3_2 represent epitope mapping of three proteins and S2_5_1, S2_5_2, S2_6_1 and S2_6_2 epitope mapping of three different proteins with K. pneumoniae antibodies, while 4_1, 4_2, 5_1 and 5_2 represent specificity assays for the first set of proteins (using C. jejuni and S. aureus antibodies) and S2_1_1, S2_1_2, S2_2_1, S2_2_2 (C. jejuni AB) as well as S2_7_1, S2_7_2, S2_8_1, S2_8_2 (S. aureus AB) for the second set of proteins.