Improved Method for Preparing N-Hydroxysuccinimide Ester-Containing Polymers for Affinity Chromatography

Abstract

N,N,N',N'-Tetramethyl(succinimido) uronium tetrafluoroborate is proposed as a reagent of choice for the activation of carboxyl groups and formation of N-hydroxysuccinimide esters on polymers. Unlike conventional methods which generate unstable gels, the reaction is appropriate for hydroxy-containing resins like Sepharose, cellulose, and dextran. The yields of activation and subsequent coupling capacity for ligands and proteins are very high. The respective columns can be used for affinity chromatography and immobilization of proteins.

Multichannel surface plasmon resonance biosensor with wavelength division multiplexing

“…C 16 alkanethiol terminated with a carboxylic head group was used to anchor antibodies, while C 11 alkanethiol chain terminated with a hydroxyl head group formed a stable non-fouling background. The carboxylic terminal groups on the sensor surface were activated with TSTU dissolved in dimethylformamide at the concentration of 1 mg/ml for 4 h [17]. After the activation, the sensor chip was rinsed with water, dried in nitrogen stream and loaded into the SPR instrument, where in situ immobilization of antibody was performed.…”

Zebrafish cyclin E regulation during early embryogenesis

“…A series of 3 injections into rabbits every 10 days of 150 pg of the purified cyclin E protein were performed. The antiserum was affinity purified on cyclin E affinity column prepared according to Wilchek (Wilchek et al, 1994).…”

A new surface plasmon resonance sensor for high-throughput screening applications

“…Sensor chips were immersed in thiol solution and stored in a dark place at room temperature for 12 h. Then, the chips were rinsed with ethanol, dried with nitrogen, rinsed with water and dried with nitrogen again. The carboxylic terminal groups on the sensor surface were activated by TSTU dissolved in dimethylformamide with concentration of 1 mg/ml for 4 h (Wilchek et al, 1994). After activation, the chips were rinsed with water, dried with nitrogen and mounted into SPR instrument.…”