Expression of single-chain antibody against RgpA protease of Porphyromonas gingivalis in Lactobacillus.
OBJECTIVE
The monoclonal antibody 61BG1.3, recognizing the RgpA protease, has been reported to confer protection against recolonization by the periodontal pathogen Porphyromonas gingivalis in humans. The aim of this study was to express a functional scFv derived from the monoclonal antibody 61BG1.3 on the surface of Lactobacillus paracasei for potential use in the prevention or treatment of periodontal diseases.


RESULTS
The scFv was fused to an E-tag and cloned in the Escherischia coli/Lactobacillus shuttle vector pLP501, which mediates surface expression of the scFv. FACS analysis using an anti-E-tag antibody revealed that the scFv was expressed on the surface of the transformed lactobacilli and binding of the scFv to RgpA was shown by ELISA. Lact. paracasei expressing the scFv against RgpA was able to agglutinate P. gingivalis whereas the Lact. paracasei expressing an irrelevant scFv fragment did not. Scanning electron microscopy demonstrated efficient binding of the lactobacilli expressing the scFv anti-RgpA to P. gingivalis.


CONCLUSIONS
We have expressed a functional scFv antibody directed against the RgpA protease of P. gingivalis in Lactobacillus.


CONCLUSIONS
These results suggest a potential of Lactobacillus expressing scFvs against P. gingivalis to be used to combat periodontal disease.
Marcotte
H
H
Department of Laboratory Medicine, Division of Clinical Immunology, Karolinska University Hospital Huddinge, Karolinska Institutet, Stockholm, Sweden. harold.marcotte@labmed.ki.se
Kõll-Klais
P
P
Hultberg
A
A
Zhao
Y
Y
Gmür
R
R
Mändar
R
R
Mikelsaar
M
M
Hammarström
L
L
D018829:Adhesins, Bacterial / Q000276:immunology*; D000372:Agglutination Tests; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000276:immunology*; D003546:Cysteine Endopeptidases / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005434:Flow Cytometry / Q000379:methods; D000080867:Gingipain Cysteine Endopeptidases; D006801:Humans; D007074:Immunoglobulin G / Q000276:immunology; D007778:Lactobacillus / Q000276:immunology*; D009055:Mouth / Q000382:microbiology; D016966:Porphyromonas gingivalis / Q000201:enzymology
D018829:Adhesins, Bacterial; D000911:Antibodies, Monoclonal; D000080867:Gingipain Cysteine Endopeptidases; D007074:Immunoglobulin G; D003546:Cysteine Endopeptidases