Affinity chromatographic purification

Most eukaryotic mRNA molecules have up to 250 adenine bases at their 3 end. These poly (A) tails can be used in the affinity chromatographic purification of mRNA from a total cellular RNA extract. Under high salt conditions, poly (A) will hybridize to oligo-dT-cellulose or poly(U)-sepharose. These materials are polymers of 10 to 20 deoxythymidine or uridine nucleotides covalently bound to a carbohydrate support. They bind mRNA containing poly (A) tails as short as 20 residues. rRNA and tRNA do not possess poly (A) sequences and will not bind. After washing the mRNA can be eluted with a low salt buffer. 

Kumar A, Bansal V, Andersson J, Roychoudhury PK, Mattiasson B (2006), Super-macroporous cryogel matrix for integrated protein isolation - immobilized metal affinity chromatographic purification of urokinase from cell culture broth of a human kidney cell line, J. Chromatogr. A 1103 35-42. 

Nevens, J. R., Mallia, A. K., Wendt, M. W., and Smith, P. K. (1992). Affinity chromatographic purification of immunoglobulin M antibodies utilizing immobilized mannan binding protein. ]. Chromatogr. 597, 247-256. 

Production, Export and Affinity Chromatographic Purification of Recombinant... 

Table 1 Comparison of the recombinant production and affinity chromatographic purification of GFP from B. megaterium [30]. Different affinity tag fusion forms of GFP were produced in II. megaterium WH323. Purification was performed using affinity chromatography. Amounts of purified GFP-Strep were determined using a Bradford protein assay kit (Bio-Rad Munich Germany) and BSA (Perbio Rockford USA) as standard. Amounts of purified GFP-His, His-TEV-GFP, Strep-Xa-GFP and Strep-TEV-GFP were calculated via their relative fluorescence per mg protein...

Fassina G, Ruvo M, Palombo G, Verdoliva A, and Marino M. Novel Ugands for the affinity-chromatographic purification of antibodies. J. Biochem. Biophys. Methods 2001 49 481M-90. 

Hematoporphyrin (3,3 -[7,12-bis-(l-hydroxyethyl)-3,8,13,17-tetramethyl-porphyrin-2,18-diyl]-dipropionic acid) [ 14459-29-1 ] M 598.7, pKest 4.8. Purified by dissolving in EtOH and adding H2O or Et20 to give deep red crystals. Also recrystd from MeOH. UV has A, ,ax 615.5, 565, 534.4 and 499.5nm in 0.1 N NaOH, and 597, 619, 634,653, 683 and 701nm in 2 N HCl. [Falk Porphyrins and Metalloporphyrins Elsevier, NY, p 175 1964.] It is used in the affinity chromatographic purification of Heme proteins [Olsen Methods Enzymol 123 324 1986.] The 0-methyl-dimethyl ester has m 203-206° (from CHCh-MeOH) and the 0,0 -dimethyl-dimethyl ester has m 145° (from CHCh-MeOH). [Paul Acta them Scand 5 389 1951.]... 

For examples of the use of latex microspheres in the affinity chromatographic purification of drug receptors, see (a) Shimizu, N., Sugimoto, K., Tang, J., Nishi,... 

TABLE 3.6 Examples of ligands suitable for affinity chromatographic purification of proteins... 

Activation of cross-linked agaroses with 1,1-carbonyl-di-imidazole yields a matrix for affinity chromatography devoid of additional charged groups/ The activated matrix (Scheme 7) is relatively stable to hydrolysis, but reacts smoothly with A-nucleophiles such as those present in affinity chromatography ligands and leashes. The matrix was treated with 1-aminobutane, 1,6-diamino-hexane, or 6-aminohexanoic acid and then with soybean trypsin inhibitor or 4-aminobenzamidine via a carbodi-imide-mediated reaction, and the products were shown to be useful for the affinity chromatographic purification of trypsin. 

Affinity chromatographic purification of an 327 arabinogalactan protein from Gladiolus... 

Nitrosulphenyl-L-methionyl)-L-tyrosyl-L-phenylalanine has been treated with aminohexyl-agarose via a carbodi-imide mediator, and the 2-nitro-sulphenyl group then removed with sodium dithionite. The matrix is suitable both for the affinity chromatographic purification of neurophysins, and for measurement of their ligand-binding parameters by quantitative affinity chromatography. 

Ace tamido-2-deoxy-D-galactose (37) Affinity chromatographic purification of a phytohaemagglutin from Robinia pseudoacacia 331 s I-... 

Acetamido-4-amino-2,4-dideoxy-D- glucose (40) Affinity chromatographic purification of a 8-D-2-acetamido-2-deoxy-glucosidase from malted barley 332 s... 

MAcetyl-L-tryptophan (36) Affinity chromatographic purification of indolyl-3-alkane a-hydroxylase 334 S o... 

Adenosine triphosphate (41) Affinity chromatographic purification of rat liver AMP-deaminase 337 ... 

Affinity chromatographic purification of pig liver phosphoglycerate kinase 338 c o to... 

Albumin, glutaraldehyde-cross-Iinked (38) Affinity chromatographic purification of hepatitis B surface antigen from human sera 340 S to... 

Alprenolol (42), (43) Affinity chromatographic purification of the /Sj-adrenergic receptor from turkey erythrocytes 341 ... 

Divinylsulphone-cross-linked agarose has been used for the affinity chromatographic purification of an agarase from a culture filtrate oi z Pseudomonas-Viikc bacterium, Agarose activated with divinylsulphone has been treated with D-mannose and the matrix used for the purification of a lectin from Onobrychis viciifolia. 

Amylose cross-linked with epichlorohydrin has been used for the affinity chromatographic purification of the extracellular isoamylase of Pseudomonas... 

Cellulose activated with epichlorohydrin has been treated with glycine and then coupled with a-amylase via a carbodi-imide mediator. Glucoamylase was also immobilized by a carbodi-imide-mediated reaction with epichlorohydrin-activated cellulose, which had previously been treated with 6-aminohexanoic acid, or by reaction with diazotized benzidine previously coupled to epichloro-hydrin-activated cellulose. Immobilized 4-chloromercuribenzoate was prepared by reaction with 1,6-diaminohexane coupled to epichlorohydrin-activated cellulose, and used for the affinity chromatographic purification of a-amylase and glucoamylase. 

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