Creating a Neutravidin chip on C1 sensor chip

Hi. I would like to create a Neutravidin chip using a C1 chip. (I have never used a C1 chip before) by NHS-EDC. Are there any suggestions on how much conc. I need to prep the Neutravidin and what buffer to use? Thanks.

Hi,
Sensor Chip C1 has a flat carboxylated surface with no dextran matrix. The
carboxyl groups support the same immobilization chemistries as the dextran
matrix on CM-series chips, but the surface of Sensor Chip C1 lacks the
hydrophilic character of the dextran matrix and the immobilization capacity is
greatly reduced (typically to about 10% of that on Sensor Chip CM5 under
comparable conditions) and the pre-concentration effect at low pH is also diminished..

(strept)avidin coupling (CM5 chip)
1. flow (5 µl/min).
2. 35 µl injection of NHS/EDC to activate the surface by modification of the carboxymethyl groups to N-Hydroxysuccinimide esters.
3. Activation of the surface has only a slight effect on the SPR signal (100–200 RU); in case of C1 may be less.
4. Injection of 35 µl (strept)avidin (150–300 µg/ml) in 20 mM acetic acid pH 4.0.
5. Deactivation of unreacted NHS-esters using 35 µl of 1 M ethanolamine (2-aminoethanol hydrochloride; Mr 97.55) pH 8.5.
6. The deactivation process with 15 µl 0.1 M H3PO4 removes any remaining electrostatically bound (strept)avidin.
7. The response of immobilized (strept)avidin is between 2000 and 6000 RU for a CM5 sensor chip.

Ref: O'Shannessy, D. J., Brigham-Burke, M. and Peck, K.; Immobilization chemistries suitable for use in the BIAcore surface plasmon resonance detector. Analytical Biochemistry (205) 1: 132-136; 1992.

Kind regards
Arnoud

Thanks Arnoud for the protocol.