A competitive enzyme-linked ligand sorbent assay (ELLSA) for quantitation of folates

An enzyme-linked ligand sorbent assay (ELLSA) for quantitation of folates is described. The method involves the following steps: (a) folate complexed to bovine serum albumin is adsorbed onto microtiter plates; (b) added folates compete with immobilized folate for binding to added biotinylated folate-binding protein; (c) biotinylated folate-binding protein bound to immobilized folate is detected after binding of avidin-alkaline phosphatase. The specificity of ELLSA is similar to that of conventional radioisotope dilution methods, and the sensitivity is high (lower limit of detection 20 fmol/sample). Quantitation of folates in erythrocyte lysates from 43 persons was performed by ELLSA. The results correlated fairly well with those obtained by the conventional radioisotope dilution method.