Detection of neu differentiation factor with a biospecific affinity sensor during chromatography

• Published in: Journal of Chromatography A Vol. 705; no. 1; pp. 163 - 169
• Main Authors: Lu, Hsieng S., Chang, David, Brankow, David, Wen, Duanzhi
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 23.06.1995
• Subjects: Animals; Biosensing Techniques; CHO Cells; Chromatography, Affinity - methods; Chromatography, Ion Exchange - methods; Cricetinae; Glycoproteins - analysis; Neuregulins; Rats; Recombinant Proteins - analysis
• ISSN: 0021-9673
• DOI: 10.1016/0021-9673(94)00737-T

A technique using a biospecific affinity sensor, BIAcore, was applied to monitor and determine mammalian cell-derived neu differentiation factor (NDF) in column fractions during chromatography. Specific purified polyclonal antibody against Escherichia coli-derived NDF was chemically bound to the surface of BIAcore sensor chips and the derivatized sensor chips were used to detect the specific binding of NDF. The measurement of NDF at very low levels can be assessed by injecting small volumes of the crude media or column fractions into the BIAcore sensor containing antibody-bound sensor chips. This automated procedure performed under computer programming control allows direct measurement of multiple NDF samples in a short period of time and provides excellent quantitative data, which is not possible using other related methods such as Western blotting, sodium dodecyl sulfate-polyacrylamide gel electrophoresis and stimulatory activity assay on receptor autophosphorylation.