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“…As indicated in Figure 1, these two peptides had different binding affinities to PK99H in the competitive ELISA assay. The apparent association constants (K,) were calculated from the 150 values as described by Nieto et al (1984). The IS0 values (peptide concentration required for 50% inhibition of PK99H binding to PAK pili) for both peptides showed that a 1,500-fold higher concentration of P A 0 peptide was required to attain the same in- …”
Section: Resultsmentioning
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“…As indicated in Figure 1, these two peptides had different binding affinities to PK99H in the competitive ELISA assay. The apparent association constants (K,) were calculated from the 150 values as described by Nieto et al (1984). The IS0 values (peptide concentration required for 50% inhibition of PK99H binding to PAK pili) for both peptides showed that a 1,500-fold higher concentration of P A 0 peptide was required to attain the same in- …”
Section: Resultsmentioning
“…If a single alaninesubstituted peptide showed a decrease in its ability to inhibit the binding of PK99H to PAK pili, the side chain Table 1. K , is the apparent association constant of monoclonal antibody PK99H for the corresponding peptide analog, which is calculated by the formula KO = l/150 (Nieto et al, 1984).…”
Section: Pk99h Epitopementioning
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“…This was done by using high (22 pg/ml) and low (0.75 pg/ml) concentrations of IgG as precoat. In theory, higheravidity RF will selectively bind to the lower concentration of IgG (antibody excess), whereas RF of all affinities will bind to the higher concentration of precoat IgG (antigen excess) (10). For these observations of RF binding avidity in the ELISA to be valid, it was critical that IgGl and IgG3 coated the microtiter plates to a similar extent, using equivalent coating concentrations.…”
Section: Resultsmentioning
“…The concentration of LPS resulting in 50% inhibition of the antibody binding was determined as described by Reed and Muench [20]. The affinity constants of monoclonal antibodies determined by this method have been shown to be in close agreement with the intrinsic affinity constant measured by Farr assay and fluorescence quenching assays [21]. Calculations of the molar concentrations were based on the assumed molecular weight of 10,000 for LPS as described by Schwartzer et al [22].…”
Section: Methodsmentioning