Objective Enzyme linked immunosorbent assay (ELISA) has been widely used for detection of antibodies or antigens, and is regarded as the gold standard in clinical diagnosis, which can provide relatively reliable, sensitive and specific results. The essence of ELISA is the specific interaction between antigens and the corresponding antibodies. However, the inherent instability of natural antibodies is the Achilles heel of ELISA, which may lead to poor reproducibility or even false results. Previously, our group created a nova gold nanoparticle-based artificial antibody, denoted as goldbody. Goldbody can specifically interact with antigens like natural antibodies, but has much better stability than that of natural antibodies. The excellent stability of goldbody makes it a potential replacement of natural antibodies in ELISA assays. Methods Herein, we demonstrate with a series of experiments that goldbody is indeed a good replacement of natural antibodies in ELISA for antigen detection. Results After necessary optimization and conjugation of horseradish peroxidase (HRP), the 13 nm HRP-labeled anti-hen egg white lysozyme (HEWL) goldbody can be used in ELISA assay to detect HEWL in the range of 1-16 mg/L. Conclusion Goldbodies can indeed replace natural antibodies in ELISA for a more accurate and consistent detection of antigens.