Protein affinity purification column

This method involves passing the protein through a column filled with resins of unique characteristics. Depending on the type of the resin or beads, purification can be achieved through (i) Ion Exchange, (ii) Size Exclusion or (iii) Affinity Chromatography. 

Purification of IgG Using Affinity Chromatography on Antigen Ligand or Protein A/G Columns... 

Fig. 3. Intein-mediated protein ligation. The IMPACT system allows affinity purification of proteins fused to an intein-CBD tag and their further isolation with a C-terminal thioester moiety (A), or an N-terminal cysteine (B). (A), N-terminal intein splicing for thioester isolation. Target protein (protein 1) is expressed in E. coli with C-terminally located intein-CBD tag. After specific binding to the chitin resin, the thiol reagent provokes the cleavage of the peptide bond between the target protein and the intein. Whereas the intein-CBD tag remains bound to the chitin resin, the protein thioester is eluted from the column. (B), C-terminal cleavage to obtain N-terminally...

Moreover, synthetic peptides can be attached to agarose beads to prepare affinity chromatography columns for the purification of receptor proteins that specifically recognize the peptides. 

The third method that is commonly used relies on the enrichment of methylated sequences. This can be achieved either by 5mC-specific antibodies or by affinity purification using protein domains that specifically bind to 5mC such as MBP or MeCP2. After purification of genomic DNA, the DNA is sheared and precipitated with a specific antibody or purified by immuno-affinity columns and can be analyzed by microarray or NGS. The same procedures can be performed by using a 5hmC-specific antibody that is also... 

Synthetic peptides can be used to isolate receptors for many hormones and other signal molec ules. For example, white blood cells are attracted to bacteria by formylmethionyl (fMet) peptides released in the breakdown of bacterial proteins. Synthetic formylmethionyl peptides have been useful in identifying the cell-surface receptor for this class of peptide. Moreover, synthetic peptides can be attached to agarose beads to prepare affinity chromatography columns for the purification of receptor proteins that specifically recognize the peptides. 

Table 1 Design Table for Screening Studies on Affinity Purification of a Protein Using Monoclonal Immunoaffinity Column (2 1 Fractional Factorial Design)...

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