Fractionation of heparin by affinity chromatography on covalently-bound human α-thrombin

1978 ◽  
Vol 83 (3) ◽  
pp. 1198-1205 ◽  
Author(s):  
Michael J. Griffith ◽  
Henry S. Kingdon ◽  
Roger L. Lundblad
Keyword(s):  
Affinity Chromatography ◽  
Covalently Bound

scholarly journals Binding of the asymmetric forms of acetylcholinesterase to heparin

1984 ◽  
Vol 221 (2) ◽  
pp. 415-422 ◽  
Author(s):  
E Brandan ◽  
N C Inestrosa
Keyword(s):  
Affinity Chromatography ◽  
Basal Lamina ◽  
Specific Binding ◽  
Heparan Sulphate ◽  
Diaphragm Muscle ◽  
Agarose Gel ◽  
Rat Diaphragm ◽  
Heparan Sulphate Proteoglycans ◽  
Sulphated Glycosaminoglycan ◽  
Covalently Bound

The interaction between acetylcholinesterase (EC 3.1.1.7) and heparin, a sulphated glycosaminoglycan, was studied by affinity chromatography. A specific binding of the asymmetric acetylcholinesterase to an agarose gel containing covalently bound heparin was demonstrated. This interaction required an intact collagenous tail, shown by the fact that the binding is abolished by pretreatment with collagenase. The globular forms did not bind to the column. Both total and intracellular asymmetric acetylcholinesterase forms isolated from the endplate region of the rat diaphragm muscle showed higher affinity for the heparin than did the enzyme from the non-endplate region. The binding to the resin was destabilized with 0.55 M-NaCl, and, among the various glycosaminoglycans tested, only heparin was able to displace the acetylcholinesterase bound to the column. Our results added further support to the concept that the asymmetric acetylcholinesterase forms are immobilized on the synaptic basal lamina via interactions with heparin-like molecules, probably related to heparan sulphate proteoglycans.

scholarly journals A simple high-yield procedure for isolation of human urinary kallikreins

1978 ◽  
Vol 171 (1) ◽  
pp. 285-288 ◽  
Author(s):  
N B Oza ◽  
J W Ryan
Keyword(s):  
Affinity Chromatography ◽  
Ph Gradient ◽  
High Yield ◽  
Antibody Preparation ◽  
Final Purification ◽  
Covalently Bound

Two human urinary kallikreins (fractions A-1 and A-2) were purified to apparently homogeneous forms. The two kallikreins were separated by affinity chromatography using Trasylol (aprotinin) covalently bound to Sepharose. The kallikreins were eluted with a pH gradient (pH 9.5-3.0). Fraction A-1 was eluted between pH 6.2 and 4.2 and fraction A-2 was eluted between pH 4.2 and 3.1. Final purification was obtained by chromatography on Sephacryl SS-200. Antibodies prepared against fraction A-2 were also reactive with fraction A-1; thus it may be possible to measure both by radioimmunoassay using the same antibody preparation.

The microstructure of functionalized poLyacrylonitrile beads for bioseparations

1990 ◽  
Vol 48 (4) ◽  
pp. 1094-1095
Author(s):  
Eduardo A. Kamenetzky ◽  
David A. Ley
Keyword(s):  
Aqueous Solution ◽  
Affinity Chromatography ◽  
Pore Size Distribution ◽  
Pore Size ◽  
Surface Coverage ◽  
Vacuum Impregnation ◽  
Thin Sections ◽  
Selective Staining ◽  
Low Viscosity ◽  
Diamond Knife

The microstructure of polyacrylonitrile (PAN) beads for affinity chromatography bioseparations was studied by TEM of stained ultramicrotomed thin-sections. Microstructural aspects such as overall pore size distribution, the distribution of pores within the beads, and surface coverage of functionalized beads affect performance properties. Stereological methods are used to quantify the internal structure of these chromatographic supports. Details of the process for making the PAN beads are given elsewhere. TEM specimens were obtained by vacuum impregnation with a low-viscosity epoxy and sectioning with a diamond knife. The beads can be observed unstained. However, different surface functionalities can be made evident by selective staining. Amide surface coverage was studied by staining in vapor of a 0.5.% RuO4 aqueous solution for 1 h. RuO4 does not stain PAN but stains, amongst many others, polymers containing an amide moiety.

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