Size exclusion chromatography recovery

Remove excess crosslinker and reaction by-products by dialysis or size exclusion chromatography. For small quantities of bait proteins, dialysis may be the better choice, because gel filtration columns often bind nonspecifically enough protein to make recoveries unacceptably low. 

Buchacher et al. [43] discussed the continuous separation of protein polymers from monomers by continuous annular size exclusion chromatography. The P-CAC used for the experiments was a laboratory P-CAC type 3 as described in Table 1. The results were compared to conventional batch column chromatography in regard to resolution, recovery, fouling, and productivity. The protein used in the studies was an IgG preparation rich in aggregates. Under the conditions used, the polymers could be separated from the monomers, although no baseline separation could be achieved in either the continuous or the batch mode. The... 

Recovery, Purity and Amino Acid Composition of CMP. The elution profiles of the CMP powders obtained on size exclusion chromatography 23) are shown in Figure 6. Both CMP isolated from WPC and whey did not contain major... 

Size exclusion chromatography (SEC, also known as gel permeation chromatography) is a method of separating compounds of different molecular masses and sizes. Because steric interactions between analytes and the stationary phase are relatively weak, unstable forms of metals can be separated from more stable complexes and from adducts stabilized by ionic interactions. Unfortunately, the process of sorption and ionic interactions between the investigated substances and the stationary phase can decrease metal recovery by as much as 50 % these interactions are also responsible for the instability of retention times [146]. The separation can be performed both in the aqueous environment and in the presence of organic solvents. Because the technique is not selective, it is utilized primarily as the first stage of multidimensional chromatography [147]. 

Chang, W.-J. Koo, Y.M. On line recovery of large molecules from mixtures using reciprocating size exclusion chromatography. Biotechnol. Tech. 1999, 13, 211-214. 

Solution phase supports have the problem of product recovery. An alternative approach is to couple the substrate to a water-soluble polymer that can be removed from solution by precipitation of the polymer or size exclusion chromatography. Water-soluble supports have been used in the enzymatic synthesis of pseudo-GM3 (118). 

X 10" to 1 X 10 in physiological solutions. On the other hand, the molecular weights of proteins are generally about 1 X 10" to 5 x 10 . Therefore, it is extremely difficult to separate LPS from protein solely by size-separation methods, such as size-exclusion chromatography (SEC) and ultrafiltration. Various procedures of LPS removal, such as ion-exchange membrane, ultrafiltration, and extraction, have been developed for pharmaproteins. These procedures, however, are unsatisfactory with respect to selectivity, adsorption capacity, and protein recovery. 

The off rate, kos, of the inhibition can be determined by calculation using Eq. (13.6) or by direct measurement. Enzyme-inhibitor complex can be isolated from excess inhibitor by size exclusion chromatography, preferably with a shift in pH to a range where the enzyme is stable but inactive, to stabilize the complex (Copp et al., 1987). It can then be added back to an activity assay, to measure the return of enzyme activity over time. The recovery of enzyme activity, koff, should be a first-order process, independent of inhibitor, enzyme, or E-I concentrations. The final rate, C, will depend on [E-I] (and any free E that might have been carried through the chromatography). 

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