p. 3
Table 1. Medium characteristics.
Mean particle size 90 μm
Particle size range 45–165 μm
Bead structure Highly cross-linked 4% agaro se, spherical
Linear flowrate
Base matrix 150–250 cm /h, 0.1 MPa (1 bar), XK 50 /60 column, bed h eight 25 cm
Swelling factor 4–5 ml drained medium/g
Coupling capacity 13–26 mg α-chymotrypsinogen/ml
pH stability*
long term 3–11
short term (CIP) 3–11
* Refers to stability of cou pling betwe en ligand abd ba se matrix. L igands can be le ss stable.

Sepharose 4 Fast Flow matrix

Sepharose 4 Fast Flow is a h ighly cross-linke d agarose matri x. In its pre-
activated CNBr form, it o ffers much improve d performance w hen compared
with the well establi shed CNBr-activa ted Sepharose 4B. T he Sepharose 4
Fast Flow matrix has hi gher rigidity an d can thus be run at high f low rates
(see Table 1).
The higher mechanical s trength of the cros s-linked matri x makes it well-
suited for use in large colu mns. Scaling up a pur ification de veloped on CNBr-
activated Sepharo se 4 Fast Flow is there fore simple and more p redictable.
The coupled product i s stable at low pH, wh ich is often require d for elution
from some immunoadsorbents.
For applications tha t require operati on at high pH, note tha t the amide bond
formed when using the co mpanion produc t NHS-activate d Sepharose 4
Fast Flow is stable up to pH 1 3 for normal use.
2. Coupling
CNBr-activated Sepha rose 4 Fast Flow is sup plied freeze-d ried in the
presence of additives . These additiv es need to be washed a way at low
pH (pH-2–3) before coupling th e desired ligand. T he use of low pH (pH2–3)
preserves the act ivity of the react ive groups, which oth erwise hydrol yse at
high pH.