COMPLEX AFFINITY PURIFICATIONS AND HIGH THROUGHPUT BUFFER EXCHANGE MADE EASY
IMCStips technology uses dispersive solid-phase extraction (dSPE) to provide simplified, highly efficient micro-purifications. The loose resin contained within IMCStips improves workflow efficiencies by facilitating the enhanced binding of analytes of interest during pipetting steps. By coupling this technology with the Dynamic Devices Lynx Series liquid handling robot, methods are streamlined with the hands-free implementation of multiple aspirate and dispense cycles that facilitate consistent results, high recoveries, and faster workflows.
BENEFITS
- Faster purification of biomolecules using various affinity resins. Complete binding of 96 samples in 1 hour!
- SizeX IMCStips process up to 96 samples in 15 minutes or less!
- Bypass challenges from other purification methods such as spin columns, magnetic beads, and fixed bed chromatography.
- Compatible with both Verified Volume Pipetting (VVP) and Standard Technology (ST) pipetting.
- Get started quickly with pre-templated scripts. We provide the scripts so you can Just Click Go!
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| Resin Options | Resin Options | Resin Options |
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Affinity IMCStips for Dynamic Devices
Increased binding across a wide range of concentrations
The results above demonstrate the automated affinity purification of polyhistidine-tagged GFP using 1 mL IMCStips on the Dynamic Devices Lynx LM1200. The IMCStips were packed with three different resin beds (25 μL, 50 μL, and 100 μL) to purify a range of polyhistidine-tagged GFP concentrations. The graphs below indicate (A-C) the percent GFP bound and (D-F) the recovery profile over 30 binding cycles for 25 μL, 50 μL, and 100 μL resin beds.
The results above demonstrate automated affinity purification of polyhistidine-tagged GFP using 5 mL IMCStips on the Dynamic Devices Lynx LM1200. (A) The binding profile of various loading concentrations of GFP over 50 mixing cycles. Higher protein amounts require an increase in binding cycles. (B) Elution volumes were tested on tips bound with 15 mg of GFP. Recovery percent plateaus at 2 mL and most of the bound fraction is recovered with 2 elutions. (C, D) Recovery of GFP from bacterial cell lysate at different GFP load amounts, using one or two elutions. Each elution used 2 mL.
SizeX IMCStips for Dynamic Devices
SizeX IMCStips® are miniaturized size exclusion chromatography columns designed for fully automated, rapid buffer exchange or desalting. SizeX IMCStips are packed with hydrophilic polymeric resin, allowing for remarkable chemical and physical stability for various size exclusion chromatography applications. SizeX IMCStips are compatible with both Verified Volume Pipetting (VVP) and Standard Technology (ST) pipetting for the Dynamic Devices Lynx® platform. Rapidly process up to 96 samples in 15 minutes or less.
SizeX IMCStips or Bio-Spin P6 columns (Spin) were used to buffer exchange 100 μL (SizeX100) or 150 μL (SizeX150) samples containing (A) 1 mg/mL of green fluorescent protein (GFP) or (B) 1 mg/mL of human IgG (IgG). The samples were processed either manually using the spin columns (Spin) or using an automated workflow on the Dynamic Devices Lynx (SizeX100, SizeX150). Data are presented as mean ± SD (n = 8). (C) Samples prepared with SizeX100 IMCStips show more consistent recoveries across various concentrations of IgG versus samples processed using spin columns. Data are presented as mean ± SD (n = 3).
*Ogata Y, Quizon PM, Nightlinger NS, Sitasuwan P, Snodgrass C, Lee LA, Meyer JD, Rogers RS. (2021). Automated multi-attribute method sample preparation using high-throughput buffer exchange tips. Rapid communications in mass spectrometry. doi: 10.1002/rcm.9222.
SizeX100 or SizeX150 IMCStips on the Dynamic Devices Lynx were used to buffer exchange 100 μL or 150 μL samples containing 9 to 158 μg of green fluorescent protein (GFP) or 8 to 149 μg of human IgG (IgG). Recoveries were measured and data are presented as mean ± SD (n = 4).
SizeX Across Multiple Sample Volumes
Protein recovery of His-tagged GFP across multiple concentrations (n=4 per concentration) measured via NanoDrop. (A) 100 μL sample volume using SizeX100 (B) 150 μL sample volume using SizeX150 (C) 750 μL sample volume using SizeX750 (D) 1000 μL sample volume using SizeX1000.
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