Assay design for a quantitative turbidimetric small hapten (700 MW)?
Coupling of a peptide via the SH of cysteine directly to the particle surface would require an amino particle. We have in the past converted CML to AML by hexanediamine (HDA) via EDAC. This can give a fully converted amino particle. then one would use SMCC (Pierce Cat. no. 22322) to link peptide sulfhydryl to particle amino. The alternative route is to prepare an albumin conjugate of the peptide using SMCC and then couple the resulting protein conjugate to a regular CML with direct EDAC protocol. For a detailed conjugation procedure see Hermanson, G.T., Bioconjugate Techniques; Academic Press1996, pp. 439 to 446. this gives detailed procedure including monitoring of the amount of incorporation of the peptide. The particle coupling may be done exactly from the Manual with direct EDAC protocol. The protein conjugate route is highly preferred. Protein conjugates of haptens are invariably more sensitive than directly coupled haptens. Controlling the colloidal stability of sensitized partic
