Required materials and reagents to begin assay development:
3.Analyte negative samples.
4.Analyte positive samples.

Step I: Reagent Preparation and Initial Assay Titration
1.Labeling of specific polyclonal antibody (with HRP, for example) for assay signal generation.
2.Selection of compatible antibodies.
3.Determination of a working titration of capture monoclonal antibody and detection polyclonal antibody conjugate.
4.Development and optimization of ELISA plate coating procedures for optimal antigen capture.

Step II: Assay Optimization
1.Selection of proper diluents for sample and conjugate signal generation.
2.Development of special additives to eliminate sample matrix effects such as interference and non-specific binding.
3.Construction of a standard curve to mimic performance characteristics of the sample matrix.
4.Development of a functional assay protocol within the target sensitivity range of the analyte.
5.Elimination or modification of extraction or sample preparation steps.

Step III: Assay Validation
1.Definition and documentation of assay performance characteristics essential for optimal assay utility (such as sensitivity and precision).
2.Documentation of diluent performance parameters to assure proper matrix composition (such as recovery and linearity).
3.Fine-tuning of specific assay components and incubation protocols to meet final performance requirements.

Step IV: Production of 25 Finished Kits
1.Quality control assessment of final components.
2.Final assembly, packaging, and delivery of components for 25 ELISA kits.

Total Assay Development Cost: range of $10,000 - $15,000
Estimated Time:
15-31 weeks
Sample Proposal 1
Monoclonal - Polyclonal Sandwich Assay Development
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