Showing papers on "Affinity chromatography published in 1970"
TL;DR: It is demonstrated that successful application of affinity chromatography in many cases will critically depend on placing the ligand at a considerable distance from the matrix backbone.
2,603 citations
TL;DR: Plasminogen was prepared from human plasma by affinity chromatography on L-lysine-substituted Sepharose with a specific activity of 100 caseinolytic units per milligram of nitrogen.
Abstract: Plasminogen was prepared from human plasma by affinity chromatography on L-lysine-substituted Sepharose. Thirty milligrams of plasminogen, with a specific activity of 100 caseinolytic units (Committee on Thrombolytic Agents) per milligram of nitrogen, were obtained from 340 milliliters of plasma. This corresponds to over 200-fold purification from plasma. Disc-gel electrophoresis at pH 8.3 indicated seven distinct bands, all of which contained activity.
2,051 citations
TL;DR: The enzyme, inactivated by an equimolar amount of mercuric chloride could be fully activated even after prolonged storage and had a specific activity about twice that of the original preparation and the Km was unchanged.
Abstract: A water insoluble derivative of a papain inhibitor was prepared by covalently linking Gly-Gly-Tyr(Bzl)-Arg to an agarose resin. The immobilized inhibitor binds active papain specifically. When papain prepared by the method of Kimmel and Smith was activated and applied to a column of the immobilized inhibitor at moderate ionic strength (20 mM EDTA, pH 4.3), about 50% of the total protein was not bound and was found to be catalytically inactive. The bound enzyme was released with distilled water. It contained one mole of SH per mole of protein. When assayed with α-N-benzoyl-l-arginine ethyl ester (25°, pH 6.0) the purified enzyme had a kcat of 28.5 sec-1 and a Km of 18 mM. The specific activity of the purified papain was about twice that of the original preparation and the Km was unchanged. The enzyme, inactivated by an equimolar amount of mercuric chloride could be fully activated even after prolonged storage.
142 citations
TL;DR: Beaded agarose (‘Sepharose’) is a useful insoluble support which has recently been used successfully in the selective purification of enzymes1–5,7,9, antibodies and antigens10–15, chemically synthesized peptides16,17, thyroxine binding serum proteins and in the study of the interaction of insulin with cell membranes.
Abstract: AFFINITY chromatography1–5, which in principle is related to the use of immunoadsorbants for the purification of antibodies6, exploits the reversible and specific interactions of ligands with macromolecules. Purification is affected by chromatographing the protein to be purified on a column containing an insoluble matrix to which the specific ligand is covalently attached. Beaded agarose (‘Sepharose’) is a useful insoluble support which has recently been used successfully in the selective purification of enzymes1–5,7,9, antibodies and antigens10–15, chemically synthesized peptides16,17, thyroxine binding serum proteins18, and in the study of the interaction of insulin with cell membranes19. In these studies the ligands or peptides were attached through their amino groups after activation of ‘Sepharose’ with cyanogen bromide20,21.
111 citations
TL;DR: A fairly simple procedure for isolating lactose synthetase A protein from human milk in 36% yield is described.
86 citations
TL;DR: A 2000-fold purification of the enzyme from bovine brain has been achieved by a combination of ammonium sulfate fractionation, DEAE-cellulose chromatography, gel filtration, and affinity chromatography; the highly purified preparation appears to be monodisperse.
58 citations
44 citations
TL;DR: The method of affinity chromatography was adapted to purification of trypsin by use of chicken ovomucoid by using the synthetic inhibitor-Sepharose resins to purify several enzymes by single step of affinity Chromatography.
44 citations
TL;DR: The present note deals with a very efficient purification procedure, based on affinity chromatography on p-aminobenzyl-I -thio-~3-D-xylopyranoside coupled, as inhibitor-ligand, to a matrix of Sepharose 2B.
40 citations
TL;DR: The same procedure was found to provide CRPs from monkey, rabbit and dog as well and may prove to be a general procedure for the isolation of CRP of other species.
35 citations
TL;DR: A highly purified RNase inhibitor from rat liver can be obtained by chromatography on columns loaded with carboxymethyl cellulose-RNase and is now available for the preparation of undegraded polyribosomes and ribosomal subunits.
TL;DR: The use of this method is illustrated by the isolation of affinity labeled peptides from staphyloccocal nuclease reacted with bromoacetyl and diazonium derivatives of deoxythymidine-3’-p-aminophenyl-phosphate-5’
Journal Article•
TL;DR: The application of these newly developed immunoadsorbents to selective removal of immunoglobulins from serum by passage on a Sepharose anti-light chain antibody column is reported.
Abstract: Currently the use of affinity chromatography has been directed toward selective purification of proteins (1). Specific antibodies have been obtained in high yield using columns of Sepharose coupled either to haptens or to protein antigens (2). It is the purpose of this communication to report the application of these newly developed immunoadsorbents to selective removal of immunoglobulins from serum by passage on a Sepharose anti-light chain antibody column. Preparation of the immunoadsorbent . Antiserum to guinea pig light chain was prepared by immunizing a goat with an IgG light chain pool, emulsified in complete Freund9s adjuvant (3). In double diffusion analysis this antiserum reacted strongly with the κ type light chains whereas the precipitin reaction against the λ type of light chains though present was significantly weaker. Five and six-tenths grams of CNBr (37.5 mg/ml of Sepharose), dissolved in water, were added with stirring at room temperature to 150 ml of packed Sepharose 4 B (Pharmacia Chemicals, Piscataway, N. J.), contained in 300 ml of 0.05 M sodium bicarbonate buffer, pH 9.0.
01 Jan 1970
TL;DR: The term affinity chromatography seems to have been first used by Cuatrecasas et al. (1968); however, the principles involved have long been known in the field of immunology.
Abstract: The term affinity chromatography seems to have been first used by Cuatrecasas et al. (1968); however, the principles involved have long been known in the field of immunology. In fact, as early as 1936, Landsteiner and van der Scheer used insolubilized haptens for the isolation of the corresponding antibodies. Since then numerous examples of antibody purification by use of immunosorbents have been reported (Silman and Katchalski, 1966).