Enrichment and immobilization of macromolecular analytes on a porous membrane utilizing permeation drag

Enrichment and immobilization of analytes by chemical bonding or physical adsorption is typically the first step in many commonly used analytical techniques. In this paper, we discuss a permeation drag based technique as an alternative approach for carrying out location-specific immobilization of macromolecular analytes. Fluorescein isothiocyanate (FITC) labeled macromolecules and their complexes were enriched near the surface of ultrafiltration membranes and detected by direct visual observation and fluorescence imaging. The level of macromolecule enrichment at the immobilization sites could be controlled by manipulating the filtration rate and thereby the magnitude of permeation drag. Higher enrichment as indicated by higher fluorescence intensity was observed at higher filtration rates. Also, larger macromolecules were more easily enriched. The feasibility of using this technique for detecting immunocomplexes was demonstrated by carrying out experiments with FITC labeled bovine serum albumin (FITC-BSA) and its corresponding antibody. This permeation drag based enrichment technique could potentially be developed further to suit a range of analytical applications involving more sophisticated detection methods. Keywords: Macromolecule, Enrichment, Immobilization, Concentration polarization, Permeation drag, Immunoassay, Membrane, Ultrafiltration