Affinity gels available

In most examples in the literature, polyclonal antibodies are used for preparing such columns but the increasing availability of monoclonal antibodies (MAbs) should lead to affinity gels based on MAbs becoming available. Such specificity would be particularly valuable where peptide drugs have to be selectively extracted from biological matrices prior to analysis. 

Chromatography suppliers offer affinity gels for a wide range of biomolecular applications. Availability... 

Earlier fluorometric methods for analysis of urinary free catecholamines have been replaced by HPLC methods that allow selective quantitation of epinephrine, norepinephrine, and dopamine. Preliminary extraction of urine is stid required and numerous preanalytical cleanup techniques are available. An alumina extraction procedure is typically coupled with ion-exchange or adsorption chromatography. Alumina pretreatment usually involves a batch extraction technique in which catechols are first adsorbed at pH 8.6 and then eluted with boric acid, which forms a complex with cis-diol groups. Purification on boric acid affinity gels provides an alternative procedure for selective adsorption of catecholamines. 

P-Galactosidase. Lysosomal P-galactosidase can be purified from human liver or placenta with conventional methods (Meisler, 1972 Sloan, 1972 Miyatake and Suzuki, 1975) or with affinity chromatography on immobilized />-aminophenyl- or 6-aminohexyl-thio-3-D-galactoside (Miller et al., 1977 Lo et al., 1979). The most rapid and convenient procedure seems to be the two-step method of Miller et al. (1977), which is reported to give a more than 20,000-fold purification with 41% yield. The affinity gel is commercially available. 

In laboratory practice, an affinity matrix (stationary phase) is tailor-made for the protein to be purified and filled into a chromatography column. For process applications many different affinity gels are commercially available. The raw extract is then passed through the column by using a physiological buffer as the mobile phase. In this process, the desired product is adsorbed selectively by the ligand and all unwanted components are washed away. 

Compared with enzymes fewer reports are available on immobilization of antibody (Ab) in sol-gels and their applications in immunosensing. Immobilization of Abs on a solid support was first reported in 1967 [128] and the technology has widespread application in affinity chromatography and other areas. However, the major problem associated with covalent immobilization of antibody on solid surface is partial loss of biological activity due to the random orientation of the asymmetric macromolecules,... 

Column chromatographic techniques were originally used as preparative tools but over the years major advances have taken place. HPLC is now a highly developed technique and a wide range of stationary phases are available. These enable partition, adsorption, gel permeation, affinity and ion-exchange chromatography to be performed. 

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