Triggering of SOSIP gp140 Envs by soluble CD4 (sCD4) was monitored via surface Plasmon resonance and was performed on a BIAcore 3000 instrument (GE Healthcare). Antibodies were immobilized using direct immobilization using amine coupling on CM3 sensor chips (GE Healthcare) at ~5000 RU. The SOSIP concentration in the presence and absence of a 1:3 mixture of sCD4 was 200 nM. Samples were injected over at a rate of 30 μl/min for a total of 90 μl using the high performance kinject injection mode with a dissociation phase of 600 s over four flow cells containing 17B, 19B, VRC01, or the Ab82 control. The chip surface was regenerated between measurements using two injections of 20 μl of glycine pH 2.0 at a flow rate of 50 μl/min. The resulting response curves were processed using the BIAevaluation 4.1 (GE Healthcare) software using a double reference subtraction. Reported response values were determined taking the average response from 170 to 175 s after the start of the injection.
Cm3 sensor chips
Manufactured by GE Healthcare
Sourced in United Kingdom
The CM3 sensor chips are a core component of GE Healthcare's analytical equipment. They are designed to monitor and measure specific parameters within a controlled laboratory environment. The CM3 sensor chips perform their function through advanced sensor technology, providing accurate and reliable data to support scientific research and analysis.
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Lab products found in correlation
Biacore 3000, by GE Healthcare (1 mentions)
Biaevaluation 4, by GE Healthcare (1 mentions)
Fluorescein isothiocyanate isomer 1 fitc, by Merck Group (1 mentions)
Sodium bicarbonate, by Merck Group (1 mentions)
Novex sharp pre stained protein standard, by Thermo Fisher Scientific (1 mentions)
Nupage mops sds running buffer, by Thermo Fisher Scientific (1 mentions)
Trizma base, by Merck Group (1 mentions)
Trizma hydrochloride, by Merck Group (1 mentions)
Vegf165, by Merck Group (1 mentions)
Instantblue, by Merck Group (1 mentions)
Sodium carbonate monohydrate, by Merck Group (1 mentions)
Dulbecco s phosphate buffered saline pbs, by Merck Group (1 mentions)
Nupage lds sample buffer 4x, by Thermo Fisher Scientific (1 mentions)
2 protocols using cm3 sensor chips
1
Monitoring SOSIP gp140 Env Triggering by sCD4
2
Characterization of Bevacizumab Binding Interactions
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Poly-ε-caprolactone (PCL; 80 kDa), 2,2,2-trifluoroethanol (TFE), basic fuchsin, Dulbecco's phosphate buffered saline (PBS), fluorescein isothiocyanate isomer I (FITC), sodium bicarbonate, sodium carbonate monohydrate, Trizma ® hydrochloride, Trizma ® base, InstantBlue and human vascular endothelial growth factor (VEGF165) were purchased from Sigma-Aldrich (UK). Bevacizumab (Avastin ® , 25 mg/mL, Genentech, US) was obtained from clinical donations that remained after an appropriate dose had been administered to patients. MicroBCA protein assays, Pierce TM Silver Stain kits and PD-10 desalting columns were procured from Thermo Fisher Scientific (UK). Novex bis-tris 4-12% SDS-PAGE gels, NuPAGE MOPS SDS running buffer, Novex Sharp pre-stained protein standard and NuPAGE LDS sample buffer (4X) were obtained from Life Technologies (UK). BIAcore consumables (including an amine coupling kit for ligand immobilisation which contained Nhydroxysuccinimide (NHS) and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), a regeneration scouting kit, and CM3 sensor chips) were sourced from GE Healthcare (UK). Vivaspin 6 ® centrifugal concentrators (molecular weight cut-off, MCWO 30 kDa) were purchased from VWR International (UK).
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