AbstractThe diffusion characteristics of several substrates of varying molecular sizes into and from Ca‐alginate gel beads in well‐stirred solutions were investigated. The values of the diffusion coefficient (D) of substrates such as glucose,L‐tryptophan, and α‐lactoalbumin [with molecular weight (MW) less than 2 × 104] into and from the gel beads agreed with those in the water system. Their substrates could diffuse freely into and from the gel beads without disturbance by the pores in the gel beads. The diffusion of their substrates into and from the gel beads was also not disturbed by increasing the Ca‐alginate concentration in the beads and the CaCl2concentration used in the gel preparation. In the case of higher molecular weight substances such as albumin (MW = 6.9 × 104), γ‐globulin (MW = 1.54 × 105) and fibrinogen (MW = 3.41 × 105), the diffusion behaviors of the substrates into and from the gel beads were very different. No diffusion of their substrates into the gel beads from solutions was observed, and only albumin was partly absorbed on the surface of the gel beads. The values ofDof their substrates from the gel beads into their solutions were smaller than their values in the water system, but all their substrates could diffuse from the gel beads. The diffusion of high molecular weight substrates was limited more strongly by the increase of Ca‐alginate concentration in the gel beads than by the increase of the CaCl2concentration used in the gel preparation. Using these results, the capacity of Ca‐alginate gel as a matrix of immobil