Affinity, generally purification

Cellobiase. Figure 2 outlines the general purification steps used to isolate a pure cellobiase (named for its function in the cellulase complex) and three forms of the hydrocellulase. In purifying cellobiase it was expedient to replace the adsorption or affinity column with a batch separation on DEAE-Sephadex A-50 and to complete the purification with cation exchange chromatography on SP-Sephadex (49, 50). Table IV is a summary of the purification of the cellobiase and co-purification of... 

A general purification method should be sufficient to purify at least a major portion of a library. Reverse-phase HPLC is generally method of choice. Affinity methods apply only to compounds with specific structural features. Nevertheless, a successful purification strategy always involves identifying the properties of the target compounds as well as those of the impurities. 

It should be emphasized that the nature of all presented protocols is very general and, thus, their application for a comprehensive characterization of your favorite multiprotein complex (YFMPC) in yeast might require only minor modifications. The logical sequence of all required steps is schematically shown in Fig. 2.1. The initial large-scale Ni affinity isolation of eIF3 followed by mass spectrometry (MS) of its subunit composition has already been described (Asano et al, 2002), and methods for identification of protein-protein interactions such as yeast two-hybrid (Y2H) and in vitro glutathione-S-transferase (GST) pull-down analysis are presented in volume 429. This chapter focuses on a description of the small-scale one-step in vivo affinity purification techniques that were used to determine the effects of deletions and... 

Puig O, Caspary F, Rigaut G, et al. The tandem affinity purification (TAP) method a general procedure of protein complex purification. Methods 2001 24 218-229. 

The detection of small molecules identified by DNA binding is based generally on the affinity purification and subsequent collection of fractions. The identification can be performed by means of the usual techniques employed for structure elucidation or quantitative determination of organic molecules... 

Table 3.19. Some forms of affinity chromatography that may be employed to purify a selected protein. Virtually all proteins carry out their biological effects by interacting in some way with other molecules, which can be given the general title ligands . This interaction is often quite biospecific. Immobilization of such ligands (or molecules which mimic ligands) onto chromatographic beads thus facilitates selective purification of the molecule of interest...

By virtue of their versatility, activated carbons (ACs) are the most frequently employed adsorbents. Since ancient times the high affinity of carbon for a wide diversity of chemical species has established it as a multi-purpose adsorbent. They are used extensively not only in daily life in domestic or office air filters but also on an industrial scale for gas purification. As they are general adsorbents they are the first choice for dealing with industrial accidents or terrorist attacks. 

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