S8

S8. mL?1 (D) and were incubated with serial dilutions of phage-displayed peptide in the presence or absence of 100 ng mL?1 benzothiostrobin. Fig. S4. Reactivity of the phage-displayed peptide N6C18 with the benzothiostrobin immunocomplex using different amounts of coating antibody. Plates were coated with antibody at 10 g mL?1 (A), 5 g mL?1 (B), 2.5 g mL?1 (C), and 1.25 g mL?1 (D) and were incubated with serial dilutions of phage-displayed peptide in the presence or absence of 100 ng mL?1 benzothiostrobin. Fig. S5. GBR 12935 DoseCresponse curves for phage clones N1C17, N2C4, N6C18. Serial dilutions of benzothiostrobin standard were mixed GBR 12935 with phage-displayed peptides in 5% methanol-PBS. Next 100 L of the mixtures were added to the antibody-coated wells. Each point represents the mean value of three replicates. Fig. S6. Effect of pH value on noncompetitive phage ELISA. Serial dilutions of benzothiostrobin standard were mixed with phage-displayed peptide in 5% methanol-PBS with different pH values, and 100 L of the mixtures were added to the antibody-coated wells. Each point represents the mean value of three replicates. Fig. S7. Effect of ionic strength on noncompetitive phage ELISA. Serial dilutions of benzothiostrobin standard were mixed with phage-displayed peptide in 5% methanol-PBS containing different ionic strengths, and 100 L of the mixtures were added to the antibody-coated wells. Each point represents the mean value of three replicates. Fig. S8. Effect of methanol on competitive phage ELISA. Serial dilutions of benzothiostrobin standard were mixed with phage-displayed peptide in PBS containing different concentrations of methanol, and 100 L of the mixtures were added to the antibody-coated wells. Each point represents the mean value of three replicates. Fig. S9. Effect of methanol on noncompetitive phage ELISA. Serial dilutions of benzothiostrobin standard were mixed with phage-displayed peptide in PBS containing different concentrations of methanol, and 100 L of the mixtures were added to the antibody-coated wells. Each point represents the mean value of three replicates. Fig. S10. Matrix interference on competitive phage ELISA. Standard inhibition curves for benzothiostrobin in the buffer, cucumber (A), tomato (B), pear (C) and rice (D) matrices using the competitive phage ELISA. Fig. S11. Matrix interference on noncompetitive phage ELISA. Standard binding curves for benzothiostrobin in the buffer, cucumber (A), tomato (B), pear (C) and rice (D) matrices using the noncompetitive phage ELISA. Fig. S12. The representative chromatograms of HPLC. Standard benzothiostrobin sample (A), blank sample of tomato (B), spiked sample of tomato (C), positive sample of tomato (D), blank sample of rice (E), spiked sample of rice (C), positive sample of rice (D). Table S1 The optimal concentrations of phage and antibody for competitive phage ELISA. Table S2 Average IC50 and Amax/IC50 ideals of the twenty competitive phage ELISAs. Table S3 Average IC50 and Amax/IC50 ideals of the competitive phage ELISA in PBS solutions GBR 12935 of various pH. Table S4 Average IC50 and Amax/IC50 ideals of the competitive phage ELISA in PBS solutions comprising different concentrations of NaCl. Table S5 Recoveries of samples spiked with benzothiostrobin by HPLC. NIHMS717694-product.docx (29M) GUID:?A3D140E1-ABF3-416B-AEEA-23F8664900B0 Abstract Twenty-three phage-displayed peptides that specifically bind to an anti-benzothiostrobin monoclonal antibody (mAb) in the absence or presence of benzothiostrobin were isolated from a cyclic 8-residue peptide phage library. Competitive and noncompetitive phage enzyme linked Mouse monoclonal to GST Tag immunosorbent assays (ELISAs) for benzothiostrobin were developed by using a clone C3-3 specific to the benzothiostrobin-free mAb and a clone N6-18 specific to the benzothiostrobin immunocomplex, respectively. Under the ideal conditions, the half maximal inhibition.