Capillary affinity chromatography

X Wu, RJ Linhardt. Capillary affinity chromatography and affinity capillary electrophoresis of heparin-binding proteins. Electrophoresis 19 2650-2653, 1998. 

ATIII has been also used as a model protein to test a novel affinity chromatographic system capillary affinity chromatography [9]. Separation quality has been found equivalent to that observed with classical affinity chromatography, whereas the necessary protein amount is strongly reduced to the nanogram level. 

To allow all culture productiou to be coutrolled, a method for rapid analysis is required. Prior to development of an LC-MS method, the analysis was both complex and time-consuming, involving the purification of a relatively large amount of the antibody using affinity chromatography, enzymatic release, and subsequent derivatizafion of the oligosaccharides and their analysis by using capillary electrophoresis. 

Amini, A., Chakraborty, A., Regnier, F.E. (2002). Simplification of complex tryptic digests for capillary electrophoresis by affinity selection of histidine-containing peptides with immobilized metal ion affinity chromatography. J. Chromatogr. B 772, 35-44. 

Cao P. and Stults J.T. (1999), Phosphopeptide analysis by on-line immobilized metal-ion affinity chromatography-capillary electrophoresis-electrospray ionization mass spectrometry, J. Chromatogr. 853(1-2), 225-235. 

Bedair, M., and El Rassl, Z. (2004). Affinity chromatography with monolithic capillary columns I. Polymethacrylate monoliths with Immobilized mannan for the separation of mannose-binding proteins by capillary electrochromatography and nano-scale liquid chromatography. /. Chromatogr. A 1044, 177-186. 

Amini, A., Pettersson, C., and Westerlund, D. (1997). Enantioresolution of disopyramide by capillary affinity electrokinetic chromatography with human alphal-acid glycoprotein (AGP) as chiral selector applying a partial filling technique. Electrophoresis 18, 950—957. 

J. D. Protein-doped monolithic silica columns for capillary liquid chromatography prepared by the sol-gel method applications to frontal affinity chromatography. Anal Chem... 

A detailed examination of the affinity of SLPI for the heparinized capillary was next made using a stepwise elution (from 0.1 to 0.9 M NaCl) (Fig. 11). SLPI eluted from the capillary with 0.2 M NaCl. This agreed well with results obtained by traditional affinity chromatography on a heparin-Sepharose matrix. The ACE method has the unique advantages over traditional affinity chromatography in that it requires much smaller quantities of protein and afforded better separation profiles. 

Total RNA or (polyA+)RNA (mRNA) can be used for experiments. In the latter case, a purification step is necessary, as mRNA is isolated from total cellular RNA by affinity chromatography on oligo-dT immobilized to a solid support. The amount of the purified RNA is determined by its dual wavelength absorbance at 260 nm and 280 nm and the quality checked by agarose gel or capillary electrophoresis. 

The majority of reports have used electrospray ionization mass spectroscopy (ESI-MS) as an analytical detection method because of its sensitivity and the soft namre of its ionization procedure, which generally only leads to the detection of the molecular ions of the positive library members. Many separation techniques have been coupled to ESI-MS, including affinity chromatography (49), size exclusion chromatography (50, 51), gel filtration (52), affinity capillary electrophoresis (53-58), capillary isoelectric focusing (59), immunoaffinity ultrafiltration (60), and immunoaffinity extraction (61). ESI-MS has also been used alone (62) to screen a small carbohydrate library. Other examples reported alternative analytical techniques such as MALDI MS, either alone (63, 64) or in conjunction with size exclusion methods (65), or HPLC coupled with immunoaffinity deletion (66). 

D. B. Kassel,T. G. Consler, M. Shallaby, P. Sekhri, N. Gordon, and T. Nadler, Direct coupling of an automated 2-dimensional microcolumn affinity chromatography-capillary HPLC system with mass spectrometry for biomolecule analysis. Techniques in Protein Chemistry VI, Academic Press, New York, 1995, pp. 39-46. 

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