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Optimization of single-antigen iELISA conditions for recombinant Brucella antigens by checkerboard titration. For each antigen, coating concentration (left), serum dilution (middle), <t>and</t> <t>HRP-conjugated</t> secondary antibody dilution (right) were optimized using standard Brucella-positive and -negative reference sera. Data points represent mean OD450 ± SD (technical triplicates). Working conditions were selected based on the maximum P/N ratio. (A) rOmp19; (B) rOmp2b; (C) rOmp31; (D) rBP26.
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Optimization of single-antigen iELISA conditions for recombinant Brucella antigens by checkerboard titration. For each antigen, coating concentration (left), serum dilution (middle), and HRP-conjugated secondary antibody dilution (right) were optimized using standard Brucella-positive and -negative reference sera. Data points represent mean OD450 ± SD (technical triplicates). Working conditions were selected based on the maximum P/N ratio. (A) rOmp19; (B) rOmp2b; (C) rOmp31; (D) rBP26.

Journal: Frontiers in Veterinary Science

Article Title: Optimizing ovine brucellosis serodiagnosis: evaluation of recombinant Brucella antigens and multi-antigen combinations for iELISA

doi: 10.3389/fvets.2026.1775573

Figure Lengend Snippet: Optimization of single-antigen iELISA conditions for recombinant Brucella antigens by checkerboard titration. For each antigen, coating concentration (left), serum dilution (middle), and HRP-conjugated secondary antibody dilution (right) were optimized using standard Brucella-positive and -negative reference sera. Data points represent mean OD450 ± SD (technical triplicates). Working conditions were selected based on the maximum P/N ratio. (A) rOmp19; (B) rOmp2b; (C) rOmp31; (D) rBP26.

Article Snippet: Optimization parameters included: (i) coating concentration of the single antigen or antigen mixture, (ii) serum dilution, and (iii) working dilution of the HRP-conjugated secondary antibody (Rabbit anti-sheep IgG (H/L): HRP; BIO-RAD, 5184–2,504).

Techniques: Recombinant, Titration, Concentration Assay