A label-free immunoassay using eggshell membrane as matrix and poly(diallyldimethylammonium chloride) as light-scattering enhancer

Abstract

A label-free immunoassay system using eggshell membrane as a matrix was developed. A common spectrofluorometer was used to collect light-scattering signals. The rabbit anti-human IgG (Ab) was first immobilized on the eggshell membrane with glutaraldehyde. Then, based on the immunoreactions and electrostatic interaction, the target human IgG antigen (Ag) and poly(diallyldimethylammonium chloride) (PDDA) were captured on the eggshell membrane. It was found that the light-scattering signal resulting from the PDDA immunotargeted on modified eggshell membrane was related to the concentration of target antigen. Under the optimal conditions, the light scattering intensity is directly proportional to the concentration of Ag in the range of 5.00-500 ng/mL (r = 0.995) with the limit of detection of 2.31 ng/mL [signal:noise ratio (S:N) = 3]. The proposed method was successfully applied to the determination of IgG in human serum, and the results were in agreement with those obtained by a general immunonephelometric method.