Peptide biosensor reagents are emerging as an alternative to typical antibody-based detection methods. Peptides can be rapidly isolated using bacterial display methods for new and emerging biothreats and can be chemic...Peptide biosensor reagents are emerging as an alternative to typical antibody-based detection methods. Peptides can be rapidly isolated using bacterial display methods for new and emerging biothreats and can be chemically synthesized for rapid, large-scale production. With the emergence of peptide biosensor reagents, there is a growing need to develop methods for characterizing binding interactions. Capillary electrophoresis (CE) is a free-solution separation method that is able to determine target and analyte binding association (Kb) and dissociation constants (Kd). In this study, the Kb, Kd, and peptide specificity of an isolated peptide binding reagent to protective antigen (PA) of Bacillus anthracis were evaluated using capillary electrophoresis at 10 and 20 kV. The relative binding specificity was rapidly assessed by measuring the peptide relative mobility shift at 20 kV at nonequilibrium using bovine serum albumin (BSA), horseradish peroxidase (HRP), and an anti-PA monoclonal antibody (mAb). The αPA peptide was shown to be highly specific for PA, with a Kd = 177 nM measured at 20 kV and Kd = 312 nM measured at 10 kV. These results show that peptides from bacterial display libraries can be rapidly tested for specificity and binding affinity, in solution, for use as a potential biosensor reagent against new and emerging biothreats.展开更多
Both the concept and the model of snug quantitative structure-activity relationship (QSAR) were proposed and developed for molecular design through constructing QSAR based on some known mode of receptor/ligand interac...Both the concept and the model of snug quantitative structure-activity relationship (QSAR) were proposed and developed for molecular design through constructing QSAR based on some known mode of receptor/ligand interactions. Many disadvantages of traditional models can be avoided by using the proposed method because the traditional models only determined upon molecular structural features in sample sets themselves. A genetic virtual screening of peptide/protein combinations (GVSPPC) is proposed for the first time by utilizing this idea to examine peptide/protein affinity activities. A genetic algorithm (GA) was developed for screening combinative targets with an interaction mode for virtual receptors. GVSPPC succeeds in disposing difficulties in rational QSAR, in order to search for the ligand/receptor interactions on conditions of unknown structures. Some bioactive oligo-/poly-peptide systems covering 58 angiotensin converting enzyme (ACE) inhibitors and 18 double site mutation residues in camel antibody protein cAb-Lys3 were investigated by GVSPPC with satisfactory results (R cu 2 > 0.91, Q cv 2 > 0.86, E RMS = 0.19?0.95), respectively, which demonstrates that GVSPPC is more interpretable in the ligand-receptor interaction than the traditional QSAR method.展开更多
文摘Peptide biosensor reagents are emerging as an alternative to typical antibody-based detection methods. Peptides can be rapidly isolated using bacterial display methods for new and emerging biothreats and can be chemically synthesized for rapid, large-scale production. With the emergence of peptide biosensor reagents, there is a growing need to develop methods for characterizing binding interactions. Capillary electrophoresis (CE) is a free-solution separation method that is able to determine target and analyte binding association (Kb) and dissociation constants (Kd). In this study, the Kb, Kd, and peptide specificity of an isolated peptide binding reagent to protective antigen (PA) of Bacillus anthracis were evaluated using capillary electrophoresis at 10 and 20 kV. The relative binding specificity was rapidly assessed by measuring the peptide relative mobility shift at 20 kV at nonequilibrium using bovine serum albumin (BSA), horseradish peroxidase (HRP), and an anti-PA monoclonal antibody (mAb). The αPA peptide was shown to be highly specific for PA, with a Kd = 177 nM measured at 20 kV and Kd = 312 nM measured at 10 kV. These results show that peptides from bacterial display libraries can be rapidly tested for specificity and binding affinity, in solution, for use as a potential biosensor reagent against new and emerging biothreats.
基金the National Chunhui Project Foundation (Grant No. 99-4-4+37)the Fok-Yingtung Educational Foundation (Grant No. 98-7-6)+5 种基金the State New Drug Project (Grant No. 1996ND1035A01)the Chongqing Municipality Applied Funda-mental Science Fund (Grant No. 01-3-6)Juche Academic Innovation Foundation for Science and Technology (Grant No. 03ZY12XT06)Chongqing University Subject Constructive Fund (Grant No. 04-10-10)State Key Laboratory for Chemobiosensors and Chemometrics under MOST Fund (Grant No. 2005012)National High Technology Research and Development (863) Program of China (Grant No. 2006AA02Z312)
文摘Both the concept and the model of snug quantitative structure-activity relationship (QSAR) were proposed and developed for molecular design through constructing QSAR based on some known mode of receptor/ligand interactions. Many disadvantages of traditional models can be avoided by using the proposed method because the traditional models only determined upon molecular structural features in sample sets themselves. A genetic virtual screening of peptide/protein combinations (GVSPPC) is proposed for the first time by utilizing this idea to examine peptide/protein affinity activities. A genetic algorithm (GA) was developed for screening combinative targets with an interaction mode for virtual receptors. GVSPPC succeeds in disposing difficulties in rational QSAR, in order to search for the ligand/receptor interactions on conditions of unknown structures. Some bioactive oligo-/poly-peptide systems covering 58 angiotensin converting enzyme (ACE) inhibitors and 18 double site mutation residues in camel antibody protein cAb-Lys3 were investigated by GVSPPC with satisfactory results (R cu 2 > 0.91, Q cv 2 > 0.86, E RMS = 0.19?0.95), respectively, which demonstrates that GVSPPC is more interpretable in the ligand-receptor interaction than the traditional QSAR method.
