Paul.xu: Created page with "Surface plasmon resonance (SPR)
For the immobilization onto CM5 chip, all antigens were
activated via thiol labeling kit (GE Healthcare, Buckinghamshire,
UK). A 0...."

2018-06-28T19:03:35Z

Created page with "Surface plasmon resonance (SPR) For the immobilization onto CM5 chip, all antigens were activated via thiol labeling kit (GE Healthcare, Buckinghamshire, UK). A 0...."

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Surface plasmon resonance (SPR) For the immobilization onto CM5 chip, all antigens were activated via thiol labeling kit (GE Healthcare, Buckinghamshire, UK). A 0.5 mg of antigens in 0.5 mL were prepared in morpholinoethanesulfonic acid (MES) buffer. Then, 0.25 mL of 2-(2-pyridinyldithio)ethaneamine (PDEA) (15 mg/mL) in 0.1 M MES buffer was added. After adding 25 mL of 0.4 M 1-ethyl-3-(3- dimethylaminopropyl)-carbodiimide (EDC), the mixtures were incubated at 25uC for 10 min. The buffers were exchanged to PBS via dialysis after the incubation. A Biacore 3000 (GE Healthcare) and CM5 chip (GE Healthcare) was used in all SPR analysis. SPR experiments were carried out at a flow rate of 10 mL/min using PBS as running buffer. For the activation of the surface of CM5 chip, 0.2 M EDC and 0.5 M N-hydroxysuccinimide (NHS) solution was injected for 2 min. And 40 mM cystamine solution was injected for 3 min, and then, 0.1 M dithioerythritol (DTE) in 0.1 sodium borate solution was injected for 3 min. After the activation of the chip surfaces, 100 mg/mL of activated antigens were injected for 7 min. For deactivation of excess reactive groups on the chip surface, 20 mM PDEA and 1 M NaCl solution in 0.1 M sodium acetate was injected for 4 min. Finally, various concentrations of isolated scFvs were injected for 5 min and the binding signal was detected. The KD values of scFvs were evaluated by measuring Kon and Koff in BIA evaluation software
[[Category:Antibodies]]

SPR - Revision history

Paul.xu: Created page with "Surface plasmon resonance (SPR)For the immobilization onto CM5 chip, all antigens wereactivated via thiol labeling kit (GE Healthcare, Buckinghamshire,UK). A 0...."

Paul.xu: Created page with "Surface plasmon resonance (SPR)
For the immobilization onto CM5 chip, all antigens were
activated via thiol labeling kit (GE Healthcare, Buckinghamshire,
UK). A 0...."