Rate-limiting mass transfer in immunosorbents: characterisation of the adsorption of paraquat-protein conjugates to anti-paraquat Sepharose 4B.
A series of paraquat-protein conjugates of different molecular size has been prepared by the coupling of paraquat hexanoate to the proteins lysozyme, ovalbumin, bovine serum albumin. The characteristics of the adsorption of these conjugates to an immunosorbent consisting of monoclonal anti-paraquat antibodies covalently immobilised to Sepharose 4B have been determined. Equilibrium adsorption isotherms were found to obey the Langmuir equation and indicated that 80% or more of the antibody binding sites were accessible to the conjugates. The rates of mass transfer of the conjugates to their adsorption sites on the immobilised antibodies was well described by a model in which mass transfer is controlled by transfer across the external film and diffusion within the porous adsorbent bead. The effective diffusivities of the conjugates within the immunosorbent were measured and has allowed the effect of the size of the adsorbing molecule on the rate of adsorption to be considered. The amount of paraquat that could be adsorbed and the rates of adsorption decreased as the size of the protein to which it is conjugated increased. The diffusivity of the conjugates within the pores of the adsorbent is reduced between two and five times compared to their diffusivities in free solution. The reduction is greater for the larger proteins and the variations of the effective diffusivities and the pore diffusivities with the molecular weight of the conjugate can be well described with simple correlations.
Horstmann
B J
BJ
Department of Chemical Engineering, University of Cambridge, UK.
Chase
H A
HA
D000818:Animals; D002417:Cattle; D007163:Immunosorbent Techniques; D007164:Immunosorbents; D051379:Mice; D009113:Muramidase; D010047:Ovalbumin; D010269:Paraquat / Q000276:immunology* / Q000302:isolation & purification*; D012685:Sepharose; D012710:Serum Albumin, Bovine
D007164:Immunosorbents; D012710:Serum Albumin, Bovine; D010047:Ovalbumin; D012685:Sepharose; D009113:Muramidase; D010269:Paraquat