Patent ID: 7514223

Claim:
A method of enriching a pool of analyte molecules with candidate analyte molecules that selectively bind a target molecule, comprising: (a) determining electrochemiluminescence assay (ECLA) responses for individual members of a pool of analyte molecules binding a target molecule; (b) applying a detection limit to the analysis of the ECLA responses, wherein an ECLA response equal to or greater than the ECLA detection limit identifies an electrochemiluminescence assay positive (ECLA + ) analyte molecule and an ECLA response less than the ECLA detection limit identifies an electrochemiluminescence assay negative (ECLA − ) analyte molecule; (c) determining immunoassay (IA) responses for individual members of the pool of analyte molecules binding the target molecule; (d) applying a detection limit to the analysis of the IA responses, wherein an IA response equal to or greater than the IA detection limit is immunoassay positive (IA + ) and an IA response less than the IA detection limit is immunoassay negative (IA − ); (e) generating a pool of candidate analyte molecules comprising: (i) immunoassay negative and electrochemiluminescence assay positive molecules (IA − /ECLA + ), and enriched for low affinity analyte molecules; (ii) immunoassay positive and electrochemiluminescence assay positive molecules (IA + /ECLA + ) or immunoassay positive and electrochemiluminescence assay negative molecules (IA + /ECLA − ), and enriched for high affinity analyte molecules; or (iii) immunoassay positive and electrochemiluminescence as say negative molecules (IA + /ECLA − ), and enriched for analyte molecules that bind the target molecule at a binding site not recognized by electrochemiluminescence assay (ECLA); and (f) confirming specific binding affinity of an analyte molecule selected from the enriched pool of candidate analyte molecules.