DETECTION OF NEU DIFFERENTIATION FACTOR WITH A BIOSPECIFIC AFFINITY SENSOR DURING CHROMATOGRAPHY

A technique using a biospecific affinity sensor, BIAcore, was applied to monitor and determine mammalian cell-derived neu differentiation fa ctor (NDF) in column fractions during chromatography. Specific purifie d polyclonal antibody against Escherichia coli-derived NDF was chemica lly bound to the surface of BIAcore sensor chips and the derivatized s ensor chips were used to detect the specific binding of NDF. The measu rement of NDF at very low levels can be assessed by injecting small vo lumes of the crude media or column fractions into the BIAcore sensor c ontaining antibody-bound sensor chips. This automated procedure perfor med under computer programming control allows direct measurement of mu ltiple NDF samples in a short period of time and provides excellent qu antitative data, which is not possible using other related methods suc h as Western blotting, sodium dodecyl sulfate-polyacrylamide gel elect rophoresis and stimulatory activity assay on receptor autophosphorylat ion.