Epitope mapping of monoclonal antibodies specific to serovar of Leptospira, using phage display technique.
Random heptapeptide library displayed by bacteriophage T7 was used to characterize epitopes of five monoclonal antibodies that were specific to L. australis, L. bangkok, and L. bratislava. Phages selected by biopanning were cloned by plaque isolation, and the binding specificity of individual clones was confirmed by enzyme-linked immunosorbent assay, before being further amplified and checked for phage peptide sequence using PCR and DNA sequencing. Almost all of the peptide epitopes were continuous or linear. Interestingly, in phages reacting with the monoclonal antibody (MAb) clones F11, F20, 2C3D4, and 8C6C4A12, the deduced amino acid sequence of the displayed peptides corresponded to a segment of hypothetical protein of the Leptospira genome (L. interrogans serovar Lai and Copenhageni). Considering the deduced amino acid sequences of phages reacting with the MAb clones F11, F20, 2C3D4, and 8C6C4A12, the consensus motif -SKSSRC-, -TLINIF-, -SSKSYR- and -CTPKKSGRC- appeared respectively. No similarity was observed among phage reacting with the MAb clone F21. The results demonstrate that T7 phage display technique has potential for epitope mapping of leptospiral MAbs, and for rapid analysis of the interactions between phage display peptides with the MAb. The finding of a phage peptide that binds to MAb with protective activity can be further tested as a candidate for leptospirosis vaccine in the future.
Ramasoota
Pongrama
P
Department of Social and Environmental Medicine, Faculty of Tropical Medicine, Mahidol University, 420/6 Rajvithi Road, Bangkok 10400, Thailand. tmprt@mahidol.ac.th
Tungtrakanpoung
Rongdej
R
Pitaksajjakul
Pannamthip
P
Ekpo
Pattama
P
Froman
Gunnar
G
Chaicumpa
Wanpen
W
D000911:Antibodies, Monoclonal / Q000276:immunology*; D000942:Antigens, Bacterial; D017123:Bacteriophage T7; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D006801:Humans; D007919:Leptospira / Q000145:classification / Q000276:immunology* / Q000302:isolation & purification; D007922:Leptospirosis / Q000175:diagnosis* / Q000276:immunology / Q000517:prevention & control; D010455:Peptides
D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D000939:Epitopes; D010455:Peptides