INVESTIGATING MEMBRANE MORPHOLOGY AND QUANTITY OF IMMOBILIZED PROTEIN FOR THE DEVELOPMENT OF LATERAL FLOW IMMUNOASSAY

Abstract

This study was aimed at gaining a quantitative understanding of the effect of protein quantity and membrane pore structure on protein immobilization. The concentration of immobilized protein was measured by staining with Ponceau S and measuring its color intensity. In this study, both membrane morphology and the quantity of deposited protein significantly influenced the quantity of protein immobilization on the membrane surface. The sharpness and intensity of the red protein spots varied depending on the membrane pore structure, indicating a dependence of protein immobilization on this factor. Membranes with smaller pores resulted in a higher color density, corresponding to enhanced protein immobilization and an increased assay sensitivity level. An increased of immobilized volume has a significant jagged outline on the protein spot but, conversely, no difference in binding capacity.

Keywords:

• immobilization
• immunoassay
• morphology
• nitrocellulose membrane
• pore size

ACKNOWLEDGMENTS

The authors wish to thank Universiti Sains Malaysia (Short Term Grant, 60311008) and Research University Grant (811178) for providing financial support for this project.