Affinity precipitation

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  • Synthetic polymers fulfill many functions in biotechnology and medicine. In cell culture technology and tissue engineering they provide the surfaces to which cells may attach. Cross-linked polymer networks are used for drug delivery and cell encapsulation. Polymer-based porous membranes can be used to shield implanted cells from the immune system of the host, while allowing for the exchange of nutrients and metabolic waste products thus keeping the cells alive and functioning.

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  • The extremely heat-stable 5¢-methylthioadenosine phos-phorylase from the hyperthermophilic archaeonPyrococcus furiosuswas cloned, expressed to high levels inEscherichia coli, and purified to homogeneity by heat precipitation and affinity chromatography. The recombinant enzyme was subjected to a kinetic analysis including initial velocity and product inhibition studies. The reaction follows an ordered Bi–Bi mechanism and phosphate binding precedes nucleo-side binding in the phosphorolytic direction....

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  • Stringent factor is a ribosome-dependent ATP:GTP pyrophosphoryl trans-ferase that synthesizes (p)ppGpp upon nutrient deprivation. It is activated by unacylated tRNA in the ribosomal amino-acyl site (A-site) but it is unclear how activation occurs. A His-tagged stringent factor was isolated by affinity-chromatography and precipitation. This procedure yielded a protein of high purity that displayed (a) a low endogenous pyrophosphoryl transferase activity that was inhibited by the antibiotic tetracycline; ...

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