Affinity column chromatography

Some members of class A high-molecular-mass PBPs were found to bind to moenomycin [64,94], E. coli PBP la and lb were purified by using moenomycin-affinity column chromatography in combination with other column chromatographic methods. 

Terasaki AG, Ohnuma M, Mabuchi I. Identification of actin-binding proteins from sea urchin eggs by F-actin affinity column chromatography. J Biochem (Tokyo) 1997 122 226-36. 

To assess the capacity and the recovery of forskolin from the affinity column, 30 /rg of forskolin was added and passed through the column (2.5 mL of gel), and the forskolin content was analyzed by ELISA. After washing with 5 column volumes of PBST, 22.5 /rg of forskolin remained bound and was then completely eluted with the PBS containing 45% of MeOH. Therefore, the capacity of affinity column chromatography was determined to be 9.4 figlmL. 

Limonoate dehydrogenase was purified from cell-free extracts of Arthrobacter globiformis by ammonium sulfate fractionation. Blue dye ligand affinity column chromatography and DEAR ion exchange HPLC (29). These steps provided a 428-fold increase in purity over the cell-free extract. The largest increase in purification... 

The enzyme was purified 178-folds with 90-95% homogeneity and showed a specific activity of 7 umol NADH oxidised min—1 mg—1 protein by emplying ion-exchange (DEAE cellulose. Mono Q), gel filteration (Sephacryl S-300) and two affinity column chromatography. The best separation of the enzyme protein was obtained with nucleotide analogue affinity ligand reactive-red 120 agarose. The enzyme... 

Z to (diH)Z have been conducted (Plant Physiol, submitted). The reaction is NADPH-dependent and does not require the presence of ATP and cations (Table 2). The enzyme has been partially purified using ammonium sulfate precipitation, anion exchange and affinity column chromatography. The affinity of the enzyme for i Ade (N -(A -isopentenyl)adenine), the unhydroxylated counterpart of Z, is negligible. Preliminary estimates of Z reductase activities varied substantially between species, with high activity in P. coccineus and P. vulgaris embryos but only marginally detectable activity in P. lunatus. 

One of the original clones secreted anti-forskolin antibody which was purified by protein A affinity column chromatography and its purity was confirmed by MALDl mass spectrometry. The MAh was bound to polystyrene microtitration plates precoated with a forskolin-human serum albumin (HAS). The full measuring range of the assay extended from 6 ng to 200 ng mP of forskolin [68, 76]. The ELISA system established in this study was more sensitive compared to TEC [47], GLC [74] or HPLC [47, 77]. 

Ogata S, Muramatsu T, Kobata A (1975) Fractionation of glycopeptides by affinity column chromatography on concanavalin A-Sepharose. J Biochem 78 687-696... 

Singh, B. R., Chai, Y. G., Robertson, D. T. and Song, P S., 1989, A photoreversible phytochrome affinity column chromatography for putative phytochrome receptor studies. J. Biochem. Biophys. Meth. 18 105-112. 

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