Fluorimetric Detection of Solid Surface - Catalysed Inactivati of Bovine Serum Albumine in Dilute Solutions

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Abstract
The time and temperature dependent inactivation of BSA in glass and polypropylene(PP) tubes are measured in dilute solutions for different electrolyte concentrations, probable adsorption mechanism is elucidated with rate constants and BSA uptake of modified poly(HEMA) membranes is investigated using fluorescence detection techniques. Initial fast and irreversible adsorption rates are found to be first order, temperature and pH sensitive, surface charge dependent and reached a maximum within 20 minutes showing a conformational change that facilitated binding. Involvement of disulfide bonds to this initial adsorption is evidenced by the reaction rates for unfolding of BSA structure at the solid surfaces which played a catalytical role. Secondary slow reversible adsorption rates are found to be first order, temperature insensitive but dependent on initial concentration, completely prevented in pre-coated tubes. Glass surfaces showed higher adsorption than the PP surfaces with 2-10 ppm BSA incubated upto an hour at 37°C. Poly(HEMA) and its negatively charged acrylic acid(AA) modified membranes show no BSA adsorption but its positively charged dimethylaminoethylmethacrylate (DMAEMA) modified membranes show significant initial uptake and 1.67 ± 0.07 ug/cm2 at the end of 24 hours. BSA uptake is proportional with the percentage of DMAEMA.