Lactobacillus helveticus MIMLh5-Specific Antibodies for Detection of S-Layer Protein in Grana Padano Protected-Designation-of-Origin Cheese |
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| Authors: | Milda Stuknyt? Eeva-Christine Brockmann Tuomas Huovinen Simone Guglielmetti Diego Mora Valentina Taverniti Stefania Arioli Ivano De Noni Urpo Lamminm?ki |
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| Institution: | aDepartment of Food, Environmental and Nutritional Sciences, University of Milan, Milan, Italy;bDepartment of Biochemistry and Food Chemistry, Division of Biotechnology, University of Turku, Turku, Finland |
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| Abstract: | 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. |
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