Immunoreaction of a recombinant nanobody from camelid single domain antibody fragment with Acinetobacter baumannii.
BACKGROUND
Acinetobacter baumannii, an important nosocominal pathogen, causes various human infections like meningitis, bacteremia and pneumonia. The aim of this study was to produce nanobodies derived from camel heavy chain antibodies (HcAb) against a conserved region of the biofilm associated protein (Bap) of A. baumannii, by phage display technique.


METHODS
A camel was immunized with the purified recombinant protein expressed from the conserved region of Bap and polyclonal antibody production was confirmed by ELISA. After RNA extraction from peripheral lymphocytes, cDNA was prepared and a phagemid library was constructed.


RESULTS
Phage particles encoding nanobodies were produced by infecting transformed cells with a M13K07 helper phage. A total of six panning rounds were performed to select high affinity clones. Screening of high affinity monoclonal nanobodies was performed using phage-ELISA. A clone with the highest absorption in monoclonal phage-ELISA was selected for soluble expression of the desired nanobody.


CONCLUSIONS
This is the first report on the expression and production of nanobodies against Bap. Increasing trends of drug resistance have shifted the focus to the role of antibodies in diagnosis and treatment of human diseases. Similarities of the produced VHH to human VH, makes the role of this nanobody promising in immunotherapy.
Department of Biology, Shahed University, Tehran, Iran.
Zahra
Z
Payandeh
Iraj
I
Rasooli
Seyed Latif
SL
Mousavi Gargari
Masoumeh
M
Rajabi Bazl
Walead
W
Ebrahimizadeh
D000151:Acinetobacter Infections / Q000276:immunology*; D040981:Acinetobacter baumannii / Q000276:immunology*; D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology*; D000915:Antibody Affinity / Q000276:immunology; D002162:Camelus / Q000276:immunology; D004195:Disease Models, Animal; D007128:Immunoglobulin Fragments / Q000276:immunology*; D011485:Protein Binding / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000276:immunology; D061905:Single-Domain Antibodies / Q000276:immunology*
D000907:Antibodies, Bacterial; D007128:Immunoglobulin Fragments; D011993:Recombinant Fusion Proteins; D061905:Single-Domain Antibodies
Acinetobacter baumannii; Biofilm associated protein; Nanobody; Phage display; VHH