Sample Proposal 4

Required materials and reagents from Client:
1.Capture antibody and detection antibody pair.
2.Purified analyte.
3.Analyte positive samples.
4.Analyte negative samples.Extraction buffer.
5.Any existing assay kits or strips for comparative studies.

Step I: Reagent Preparation
1.Purification of analyte-specific monoclonal or polyclonal IgG.
2.Affinity purification of analyte-specific polyclonal IgG.
3.Conjugation of analyte-specific monoclonal IgG (if available) or polyclonal IgG to colloidal gold.
4.Preparation of blocking buffers.
5.Preparation of conjugate diluents.
6.Preparation of extraction buffers.

Step II: Selection of Materials and Assay Optimization
1.Selection of solid step membrane for best lateral flow results.
2.Selection of absorbent media for proper sample handling and flow-rate.
3.Selection of conjugate release media for efficient release of colloidal gold-antibody conjugates.
4.Selection of proper backing material for required rigidity.
5.Titration of capture antibodies and colloidal gold-antibody conjugates to obtain required sensitivity levels for each analyte.
6.Determination of proper blocking buffer to eliminate nonspecific background signals.
7.Development of user-friendly test protocol for field use.

Step III: Assay Validation
1.Definition and documentation of assay performance characteristics essential for optimal assay utility (such as sensitivity and precision).
2.Comparison of results to any current standard ELISA results.
3.Provide 100 test units to client for evaluation.

Total Cost: range of $10,000 - $15,000
Total Time: 8-14 weeks

Required materials and reagents from Client:
1.Several different affinity purified PoAb.
2.Purified natural analyte.
3.Purchased analyte for use in colloidal gold conjugations.
4.Positive samples.
5.Negative samples.
6.Any current tests for comparison.

Step I: Feasibility Study, Reagent Preparation
Each polyclonal will be assessed for its ability to bind analyte-colloidal gold conjugates. Upon completion of Step I, all reagents will be prepared.
1.Conjugation of the analyte to colloidal gold in 3 different sizes (3-6 nm, 8-12 nm, and 17-23 nm gold particles).
2.Preparation of blocking buffers.
3.Preparation of conjugate diluents.
4.Assessment of each PoAb to bind the analyte-colloidal gold.

Step II: Feasibility Study, Selection of Rapid Test Materials and Assay Optimization
All antibodies that successfully bind the analyte-colloidal gold conjugate will be used during this step. The antibodies will be examined individually and in combinations. Step III will begin when the required sensitivity level is reached.
1.Selection of solid phase membrane for best lateral flow results.
2.Selection of absorbent media for proper sample handling and flow-rate.
3.Selection of conjugate release media for efficient release of conjugate
4.Selection of proper backing material for required rigidity.
5.Titration of capture antibodies and analyte-colloidal gold conjugates to obtain required sensitivity levels.
6.Selection of proper blocking buffer to eliminate nonspecific background signals.
7.Development of a user-friendly test protocol for field use.

Step III: Assay Optimization and Validation
Upon completion of Step III, the assay and assay protocol will be complete. All documentation will be in place for manufacturing and the assay will be in its final form.
1.Definition and documentation of assay performance characteristics essential for optimal assay utility (such as sensitivity and precision).
2.Determination of cross-reactivity to other molecules, as recommended by the client.
3.Comparison of results to existing test methods.
4.Final adjustments to the assay protocol.

Step IV: Production and Delivery of 10,000 Tests
1.Begin pilot scale manufacturing.
2.Quality control assessment of final components.
3.Final assembly, packaging, and delivery of components for 10,000 rapid test kits.

Total Cost: range of $10,000 - $15,000
Total Time: 15-24 weeks