Journal: Scientific Reports

Article Title: Pyruvate dehydrogenase complex—enzyme 2, a new target for Listeria spp. detection identified using combined phage display technologies

doi: 10.1038/s41598-020-72159-4

Figure Lengend Snippet: Identification of the target via MS and ORFeome phage display. ( A ) SDS-PAGE and immunoblot after IMS with GSM133-A4 scFv-Fc; M is the protein ladder Precision Plus Protein Unstained (Bio-Rad). ( B ) Alignment of the first 209 amino acids of a reference PDC-E2 sequence (WP_107899613.1) with the MSR of scFv-Fc GSM130-H1, GSM133-A4, and GSM133-E2 after ORFeome phage display. ( C ) Titration curves of the 4 initial scFv-Fc [GSM133-A4 (blue), GSM130-H1 (purple), GSM133-E2 (green), and GSM134-C1 (yellow)] against different amounts of monoclonal phage isolated in the ORFeome phage display for target identification. The setup of the assay is represented below the graphic. The complete SDS-PAGE and immunoblot images can be found, respectively, as Supplementary Figs and .

Article Snippet: Figure 2 Identification of the target via MS and ORFeome phage display. ( A ) SDS-PAGE and immunoblot after IMS with GSM133-A4 scFv-Fc; M is the protein ladder Precision Plus Protein Unstained (Bio-Rad). ( B ) Alignment of the first 209 amino acids of a reference PDC-E2 sequence (WP_107899613.1) with the MSR of scFv-Fc GSM130-H1, GSM133-A4, and GSM133-E2 after ORFeome phage display. ( C ) Titration curves of the 4 initial scFv-Fc [GSM133-A4 (blue), GSM130-H1 (purple), GSM133-E2 (green), and GSM134-C1 (yellow)] against different amounts of monoclonal phage isolated in the ORFeome phage display for target identification.

Techniques: SDS Page, Western Blot, Sequencing, Titration, Isolation, Drug discovery

Journal: Scientific Reports

Article Title: Pyruvate dehydrogenase complex—enzyme 2, a new target for Listeria spp. detection identified using combined phage display technologies

doi: 10.1038/s41598-020-72159-4

Figure Lengend Snippet: Test of GSM133-A4 in immunoblot of Listeria protein fractions and immunofluorescence microscopy. ( A ) Immunoblot of GSM133-A4 of protein fractions from the cytoplasm and cell wall of L. innocua DSM20649 (Linn), as well as E. coli BLR (DE3) extract used as a negative control. The blot shows that GSM133-A4 antibody binds to a target contained both in the cytoplasm and cell wall fractions of Listeria . The used protein ladder was Precision Plus Protein All Blue (Bio-Rad). ( B ) Fluorescence microscopy on alive L. monocytogenes 4b DSM 15675. The signal of GSM133-A4 (c and f) showed to be higher than both the negative control with a non-related antibody (a and d) and the anti-InlA positive control (b and e). The complete immunoblot image can be found as Supplementary Fig .

Article Snippet: Figure 2 Identification of the target via MS and ORFeome phage display. ( A ) SDS-PAGE and immunoblot after IMS with GSM133-A4 scFv-Fc; M is the protein ladder Precision Plus Protein Unstained (Bio-Rad). ( B ) Alignment of the first 209 amino acids of a reference PDC-E2 sequence (WP_107899613.1) with the MSR of scFv-Fc GSM130-H1, GSM133-A4, and GSM133-E2 after ORFeome phage display. ( C ) Titration curves of the 4 initial scFv-Fc [GSM133-A4 (blue), GSM130-H1 (purple), GSM133-E2 (green), and GSM134-C1 (yellow)] against different amounts of monoclonal phage isolated in the ORFeome phage display for target identification.

Techniques: Western Blot, Immunofluorescence, Microscopy, Negative Control, Fluorescence, Positive Control