Membrane adsorption of β2-microglobulin: Equilibrium and kinetic characterization

William R. Clark, William L. Macias, Bruce A. Molitoris, N. H.Linda Wang

Research output: Contribution to journalArticle

54 Scopus citations

Abstract

Enhanced extracorporeal removal of β2-microglobulin (β2m) may prevent the developmene of dialysis-related amyloidosis (DRA). One mechanism of β2m removal is membrane adsorption. Therefore, we fundamentally characterized β2m adsorption to the highly permeable polyacrylonitrile (PAN) membrane. Porous and nonporous PAN fragments were incubated in buffer containing 125I-β2m. Over a concentration range of 8 to 60 mg/liter, the equilibrium adsorption isotherm was linear (r = 0.99) for porous PAN while the isotherm for nonporous PAN suggested either multilayer binding or adsorption of proteins with differing orientations. In kinetic analyses, the approach to equilibrium versus (time) 1/2 was evaluated. For both porous and nonporous PAN, this relationship was linear (r = 0.99), consistent with a diffusion-controlled process. Adsorption reversibility was assessed by comparing the amount bound at varying residence times (0 to 4 hr) to the amount remaining adsorbed after a subsequent incubation in buffer. The fractions remaining bound at 60, 120, and 240 minutes (0.34 ± 0.02, 0.36 ± 0.06, and 0.44 ± 0.03; mean ± SEM) were significantly greater (P < 0.05) than the value at five minutes (0.23 ± 0.01). This suggests membrane-induced conformational changes in adsorbed β2m. This investigation permits the comparison of β2m adsorptive properties of PAN to those of other membrane-based and nonmembrane-based therapies designed to prevent DRA.

Original languageEnglish (US)
Pages (from-to)1140-1146
Number of pages7
JournalKidney international
Volume46
Issue number4
DOIs
StatePublished - Oct 1994
Externally publishedYes

ASJC Scopus subject areas

  • Nephrology

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