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[…] ary Material). Preliminary ELISA results indicated that the two peptides did not cross-react with the SV40 and JCPyV hyperimmune sera employed (see data in the Section “”). The amino acid (a.a.) sequences of the two peptides, known as VP1-L and VP1-M, which start at the N-terminal with a.a. L and M, respectively, are as follows: VP1-L: NH2—LKLSAENDFSSDSPERK—COOHVP1-M: NH2—MLNLHAGSQKVHEHGGGK—COOH, VP1-L: NH2—LKLSAENDFSSDSPERK—COOH, VP1-M: NH2—MLNLHAGSQKVHEHGGGK—COOH, Amino acid sequences of BKPyV viral capsid protein 1 (VP1), named peptide L viral capsid protein 1 (VP1 L) and VP1 peptide M (VP1 M), were characterized regarding stable secondary structure formation. Analysis was carried out by PSIPRED server (–). Moreover, peptide sequences were mapped on native virion proteins to verify structural similarities. VP1-L and VP1-M, in their monomeric forms, were obtained from computational prediction carried out by I-TASSER server (–). Molecular visualizations were performed by PyMOL (PyMOL Molecular Graphics System, Version 1.3, Schrödinger, LLC). Computational tools were available through ExPASy server ()., Indirect ELISA was developed and standardized to detect specific antibodies against BKPyV VP1 in human sera using VP1-L and VP1-M synthetic peptides., Plates were coated with 5 μg of the selected peptide for each well and diluted in 100 μl of Coating Buffer (Candor Bioscience, Wangen im Allgäu, Germany) at 4°C for 16 h., Blocking was made with 200 μl/well of the Blocking Solution (Candor Bioscience, Germany) at 37°C for 90 min., Wells were covered with 100 μl of serum sample diluted 1/20 in low cross-buffer (Candor Bioscience, Germany). Different wells were co […]

Pipeline specifications

Software tools PSIPRED, I-TASSER, PyMOL