CEInfinite iCIEF Solutions

CEInfinite AESlytes Carrier Ampholytes

Carrier ampholytes are essential for isoelectric focusing (IEF) separation of proteins. The fundamental requirements of good Carrier ampholytes for IEF applications are good pH linearity, high buffering capacity, small ΔpK (less than 4), good stability, and insignificant influence on the sample. 

Since early Carrier ampholytes were developed before the introduction of imaged capillary IEF (iCIEF), the unique requirement of low UV absorbance could not be applied in iCIEF application. In addition, interaction of Carrier ampholytes with protein in gel media and solution phase is different. Therefore, it is advantageous to have Carrier ampholytes for iCIEF with low protein interaction in solution phase for reproducible iCIEF protein separation and with low UV absorbance for minimised electropherogram baseline noise.

The AESlytes Carrier ampholytes were developed specifically for protein high performance iCIEF separation based on our extensive research.

In addition to meet all the requirements of good Carrier ampholytes, such as 

  • good pH linearity
  • high buffering capacity
  • good stability
  • low protein interaction
  • low UV absorbance

AESlytes are composed of a remarkably high percentage of effective Carrier ampholytes (CAs) with small ΔpK, which provides faster focusing and more stable pH gradient. Proteins are so complex, and they react differently with CAs, depending on the functional groups of CAs. Therefore, we developed three series of AESlytes that possess different basic functional groups to minimise protein interaction. AESlytes are available in High Resolution (HR), Super High (SH) and Ultra High (UH) resolution series.

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CEInfinite AESlytes use for iCIEF of Monoclonal Antibodies (mAbs)

Both HR and SH CEInfinite AESlytes are good for iCIEF of monoclonal antibodies (mAbs) (see below figure), fusion proteins, antibody drug conjugates (ADCs), dairy proteins, meat proteins and seed proteins.

CEInfinite AESlytes use for iCIEF of Acidic Fusion Proteins

SH CEInfinite AESlytes are especially good for iCIEF of acidic fusion proteins (see below figure), dairy proteins, meat proteins and seed proteins; while HR CEInfinite AESlytes are notably good for neutral and basic fusion proteins.

CEInfinite AESlytes use for improving Separation Efficiency of some ADCs

The separation efficiency of some ADCs is improved when mixing SH and HR CEInfinite AESlytes (see below figure).

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CEInfinite AESlytes available for Fusion Proteins and Other Complex Proteins

UH CEInfinite AESlytes are specially designed for separation of fusion proteins and other complex proteins

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CEInfinite AESlytes available in Broad Range and Narrow Range pH Intervals

CEInfinite AESlytes (HR, SH and UH) are provided at broad-range and narrow range pH intervals. Broad range CEInfinite AESlytes allow for a universal iCIEF method for a range of proteins with different isoelectric points and delivers fast focusing of proteins. Narrow range CEInfinite AESlytes provide higher separation efficiency at the cost of slower focusing of proteins (see below figure). Specific iCIEF methods have to be developed when the pI of proteins is in different ranges.

CEInfinite AESlytes versus Vendor CAs

CEInfinite AESlytes have demonstrated superior focusing and low UV noise. The below figure illustrated focusing of 6 pI markers with CEInfinite AESlyte SH pH 4-8 and pH 3-7 from another vendor. Much higher baseline noise was observed from the electropherogram with CAs from the other vendor. Furthermore the pI 5.85 did not complete focusing with CAs from the other vendor under the same experimental conditions.

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CEInfinite AESlytes for iCIEF-MS

CEInfinite AESlytes allow direct coupling of iCIEF to a mass spectrometer (MS). CEInfinite AESlytes are developed to be MS compatible (contain low salt) and have an extremely high percentage of good amphoteric components. 

High protein iCIEF resolution can be achieved even with very low AESlyte concentration (0.5%). These advantages greatly minimise possible interference of MS signal by salts and CAs. 

In addition, “widened” protein peaks with narrow range CAs help with mobilisation of focused protein peaks into the transfer capillary, further reducing protein remixing.

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Offline iCIEF-MS

The CEInfinite Preparative instrument allows you also to perform offline iCIEF-MS by protein fractionation. The collected fractions can be further analysed with MS.

Overview of the CEInfinite AESlytes Carrier Ampholytes

We promise our researchers to provide them consistent quantity and quality of AESlytes CAs. 

CEInfinite AESlytes CAs are synthesised strictly to ensure a high performance iCIEF separation. Due to their low interaction with proteins increases resolving power and CEInfinite AESlytes CAs do not strongly absorb UV and eliminating baseline noise. Finally, their linear pH gradient makes for an outstanding accurate pI calibration.

Overview of other CEInfinite Solutions

All CEInfinite test kits, solutions are fully tested and certified. The products are manufactured under strict quality control guidelines to save you time as well as eliminating variability for a more reproducible result.

Please note that the CEInfinite AESlytes CAs and other solutions can be used for any iCIEF as well as for CIEF assays. 

If you need any additional information, please do not hesitate to contact us.

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Custom Made Solution Services

We understand the complexity of a protein. To achieve higher resolution and better separation of your current (i)CIEF assays, we also offer custom made AESlytes CAs upon request. Please contact us for further support on this custom-made solution services.

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