Isolation of scFv fragments specific to OmpD of Salmonella Typhimurium.
Pork meat is one of the major sources for human infections with Salmonella enterica subspecies enterica serovars. Further, zoonoses caused by S. enterica subspecies enterica serovars are responsible for substantial economical losses in industrial countries. Quick and reliable detection of this infection is urgently needed to improve consumer security. Due to its capability to identify infections independent of the species, a competitive ELISA is the preferable method for the detection of anti-Salmonella antibodies in serum. Recombinant antibody fragments (scFvs) were isolated from the naive human antibody gene library HAL7 by phage display. Recombinant produced outer membrane protein D (OmpD) of Salmonella Typhimurium was used as antigen. The characterization of the isolated single chain Fv (scFv) antibodies was done by enzyme-linked immunosorbent assay (ELISA), immunoblot, sequencing, epitope mapping and size exclusion chromatography (SEC). The detection of anti-OmpD IgGs in swine sera by competitive ELISA was shown in a proof of principle concept. Furthermore, the developed competitive ELISA would be compatible to a recently published DIVA vaccine, allow to distinguish between infected and vaccinated pigs.
Meyer
Torsten
T
Technische Universität Braunschweig, Institut für Biochemie und Biotechnologie, Abteilung Biotechnologie, Spielmannstr. 7, 38106 Braunschweig, Germany.
Stratmann-Selke
Janin
J
Meens
Jochen
J
Schirrmann
Thomas
T
Gerlach
Gerald F
GF
Frank
Ronald
R
Dübel
Stefan
S
Strutzberg-Minder
Katrin
K
Hust
Michael
M
D000818:Animals; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D018604:Epitope Mapping; D005516:Food Microbiology / Q000379:methods; D015723:Gene Library; D018272:Porins / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology / Q000302:isolation & purification; D012480:Salmonella Infections / Q000175:diagnosis; D012486:Salmonella typhimurium / Q000276:immunology*; D012680:Sensitivity and Specificity; D057127:Single-Chain Antibodies / Q000276:immunology / Q000302:isolation & purification*; D013552:Swine; D013553:Swine Diseases / Q000175:diagnosis
C470933:OmpD protein, Salmonella typhimurium; D018272:Porins; D011994:Recombinant Proteins; D057127:Single-Chain Antibodies