A Protocol for the Optimization of Lateral Flow Immunoassay Strip Development

Abstract

Background: Lateral flow immunoassay (LFIA) is a well-known method because of its extensive application in many fields, such as medical, food safety, and environmental analysis. LFIA is also under development for pathogen or protein detection, where rapid detection is essential. However, there are few studies on the optimal process for LFIA production. Methods: This study aimed to evaluate the roles of the different materials that make up the LFIA strip, consisting of the sample pad, conjugate pad, nitrocellulose membrane, and absorbent pad. Different membranes and pads were evaluate regarding membrane permeability, generation of the test line, capillary flow time, and result visibility. Results: The results indicated that the optimized LFIA strip is composed of four membranes and pads, including the CF3 sample pad, FF170HP nitrocellulose membrane immobilized with 1 µg of antibody, Standard 17 conjugate pad, and CF6 absorbent pad. The result was visually detectable within 15 minutes. Conclusion: This optimization protocol could be a reference for the development of new test strips.