Lactobacillus helveticus MIMLh5-specific antibodies for detection of S-layer protein in Grana Padano protected-designation-of-origin cheese.
Single-chain variable-fragment antibodies (scFvs) have considerable potential in immunological detection and localization of bacterial surface structures. In this study, synthetic phage-displayed antibody libraries were used to select scFvs against immunologically active S-layer protein of Lactobacillus helveticus MIMLh5. After three rounds of panning, five relevant phage clones were obtained, of which four were specific for the S-layer protein of L. helveticus MIMLh5 and one was also capable of binding to the S-layer protein of L. helveticus ATCC 15009. All five anti-S-layer scFvs were expressed in Escherichia coli XL1-Blue, and their specificity profiles were characterized by Western blotting. The anti-S-layer scFv PolyH4, with the highest specificity for the S-layer protein of L. helveticus MIMLh5, was used to detect the S-layer protein in Grana Padano protected-designation-of-origin (PDO) cheese extracts by Western blotting. These results showed promising applications of this monoclonal antibody for the detection of immunomodulatory S-layer protein in dairy (and dairy-based) foods.
Department of Food, Environmental and Nutritional Sciences, University of Milan, Milan, Italy.
Milda
M
Stuknyte
Eeva-Christine
EC
Brockmann
Tuomas
T
Huovinen
Simone
S
Guglielmetti
Diego
D
Mora
Valentina
V
Taverniti
Stefania
S
Arioli
Ivano
I
De Noni
Urpo
U
Lamminmäki
D000911:Antibodies, Monoclonal / Q000276:immunology*; D001435:Bacteriophages / Q000276:immunology; D015153:Blotting, Western; D002611:Cheese / Q000382:microbiology*; D004926:Escherichia coli / Q000235:genetics; D005504:Food Analysis / Q000379:methods*; D006801:Humans; D052198:Lactobacillus helveticus / Q000276:immunology*; D008562:Membrane Glycoproteins / Q000276:immunology* / Q000378:metabolism; D019151:Peptide Library; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D057127:Single-Chain Antibodies / Q000235:genetics
D000911:Antibodies, Monoclonal; D008562:Membrane Glycoproteins; D019151:Peptide Library; D011994:Recombinant Proteins; C055764:S-layer proteins; D057127:Single-Chain Antibodies