Potential of dual-skinned, high-flux membranes to reduce backtransport in hemodialysis.

BACKGROUND: Potential backfiltration of cytokine-inducing material is a clinical concern during hemodialysis conducted with high-flux membranes. Novel hollow-fiber membranes were developed that had asymmetric convective solute transport properties, aimed at reducing the passage of potentially harmful molecules from dialysate to blood, while maintaining the desired fluid and solute movement from blood to dialysate. METHODS: Sieving coefficient as a function of molecular weight was measured in vitro using polydisperse dextrans. Measurements were conducted using two different flat-sheet membranes in series or using hollow fiber membranes having two integrally formed skin layers. Based on measured experimental parameters, model calculations simulated the performance of a clinical-scale dialyzer containing these new membranes versus that of a commercially available high-flux dialyzer. RESULTS: Asymmetric convective solute transport was demonstrated using both commercial flat-sheet and newly developed hollow-fiber membranes. For two flat-sheet membranes in series, the extent of asymmetric transport was dependent on the order in which the solution was filtered through the membranes. For the hollow-fiber membranes, the nominal molecular weight cut-off was 20 kD in the blood-to-dialysate direction and 13 kD in the dialysate-to-blood direction. For this membrane, model calculations predict that clearance of a beta2-microglobulin-sized molecule (11,800 D) would be significantly greater from blood to dialysate than in the reverse direction, even under conditions of zero net ultrafiltration. CONCLUSION: A novel hollow-fiber dialysis membrane was developed that allows greater convective solute transport from blood to dialysate than from dialysate to blood.

Equilibrium adsorption of LDL and gold immunoconjugates to affinity membranes containing PEG spacers.

The objective of this study was to provide insight into the effects of spacer chemistry on immunoaffinity separations for the capture of large macromolecules and biological complexes. Immunoaffinity membranes were prepared by immobilization of immunoglobulin G (IgG) to flat sheet microporous membranes. Two different systems were examined: immobilized IgG for the immunoadsorption of human low-density lipoprotein (LDL) and immobilized IgG for the immunoadsorption of gold particle immunoconjugate. The IgG was immobilized either directly to the membrane or via a polyethylene glycol (PEG) spacer. Adsorption of LDL was significantly greater for anti-LDL IgG immobilized via PEG than for IgG immobilized directly to the membrane. With the PEG spacer, the adsorption capacity for LDL matched the theoretical density of a monolayer of LDL particles on the membrane surface. The gold particle immunoconjugate, similar in size to LDL, was examined as a generalized model of restrictions to immunoaffinity adsorption of large (>20 nm) biological complexes. Adsorption of gold particles was greater for IgG immobilized via PEG than for IgG immobilized directly to the membrane. It is postulated that the PEG spacer allows lateral movement of the immobilized IgG and dense monolayer packing of adsorbed particles on the membrane surface. These results are pertinent to the removal of LDL from human plasma and the purification of gene therapy delivery vectors, viral vaccines, and other large biological complexes.

In vitro characterization of a membrane-based low-density lipoprotein affinity adsorption device.

The objective of this study was to explore the use of microporous membranes as an alternative substrate to porous beads in affinity adsorption of low-density lipoprotein (LDL) for therapeutic purposes. Flat sheet immunoaffinity membranes containing a polyclonal antibody preparation were utilized as the affinity substrate. The antibody was covalently immobilized to the surface through a poly(ethylene glycol) (PEG) spacer. Equilibrium adsorption of LDL from plasma was measured. Adsorption from plasma and elution of bound LDL using citrate buffer were studied as a function of flow rate. Specific capacity was as great as 2 mg apolipoprotein B per milliliter membrane volume. The superior transport properties of the membrane allowed rapid adsorption and regeneration, which translated to a large number of adsorptive cycles that can be performed within a given treatment time. On the basis of in vitro performance characteristics, it is estimated that an immunoaffinity membrane device can provide a reduction in patient plasma LDL concentration comparable to that provided by packed columns, but with almost an 80% reduction in the device volume.

Oral treatment for jaundice using immobilized bilirubin oxidase.

Jaundice is characterized by an excessive accumulation of bilirubin in the blood and tissues. A novel approach to reduce plasma levels of bilirubin by blocking its enterohepatic circulation was investigated. The treatment consisted of oral administration of immobilized bilirubin oxidase, which could oxidize bilirubin in the intestine to less toxic and more water-soluble products. In vivo administration of 0.1 to 2.0 mg/day of immobilized enzyme over a four-day period to chronically jaundiced Gunn rats effectively lowered plasma bilirubin levels, but only when the molar ratio of total serum bilirubin to rat serum albumin (B/RSA) was larger than 0.35. Plasma bilirubin concentration decreased in that group from an initial value of 11.3 to 6.3 mg/dl (-40%, n = 5) after eight days. This decrease was statistically significant (p < 0.05 by Student's t test). However, administration of bilirubin oxidase to rats with a B/RSA ratio less than 0.35 (n = 10) resulted in no statistically significant change in plasma bilirubin concentration.