US 4,507,229 A · Tissue protein PP.sub.4, a process for isolating it and its use

Abstract

The protein PP.sub.4 which has (a) an electrophoretic mobility in the range between that of .alpha..sub.1 and .alpha..sub.2 globulins; (b) an isoelectric point of 4.85.+-.0.15; (c) a sedimentation coefficient s.sub.20,w.sup.0 of 3.3.+-.0.2S; (d) a molecular weight determined in polyacrylamide gel containing sodium dodecylsulfate (DSD) of 35,000.+-.5,000; (e) an extinction coefficient E.sub.1 cm.sup.1% (280 nm) of 5.9.+-.0.6; (f) a carbohydrate content of 2.4.+-.0.94% (g/100 g) (mannose 0.3.+-.0.2%, galactose 0.4.+-.0.2%, xylose 0.1.+-.0.4%, glucose 0.2.+-.0.1%, glucosamine 1.0.+-.0.2% and neuraminic acid 0.4.+-.0.2%) and (g) a specified aminoacid composition, and a process for its preparation are described. PP.sub.4 can be used to prepare antisera which can be employed to detect and determine PP.sub.4 in body fluids in order to diagnose diseases of particular organs, as a "marker" to monitor the progress of a disease or to check therapy.

Claims4

Dependent claims

Protein PP.sub.4, which has the following characteristics

(a) an electrophoretic mobility in the range between that of .alpha..sub.1 and .alpha..sub.2 globulins;
(b) an isoelectric point of 4.85.+-.0.15;
(c) a sedimentation coefficient s.sub.20,w.sup.O of 3.3.+-.0.2S;
(d) a molecular weight determined in polyacrylamide gel containing sodium dodecylsulfate (DSD) of 35,000.+-.5,000;
(e) an extinction coefficient E.sub.1 cm.sup.1% (280 nm) of 5.9.+-.0.6;
(f) a carbohydrate content of 2.4.+-.0.94% (g/100 g) (mannose 0.3.+-.0.2%, galactose 0.4.+-.0.2%, xylose 0.1.+-.0.04%, glucose 0.2.+-.0.1%, glucosamine 1.0.+-.0.2% and neuraminic acid 0.4.+-.0.2%) and
(g) the following aminoacid composition:
______________________________________ Residues per 100 residues Coefficient of Aminoacid (mol-%) variation ______________________________________ lysine 6.95 1.14 histidine 0.97 17.4 arginine 5.44 1.77 aspartic acid 11.41 1.68 threonine 6.78 2.40 serine 6.21 2.26 glutamic acid 12.25 0.43 proline 1.96 6.20 glycine 6.68 3.83 alanine 7.92 1.67 cystine 1/2 0.77 19.5 valine 5.34 3.80 methionine 1.98 6.00 isoleucine 5.21 2.23 leucine 11.50 0.45 tyrosine 3.55 4.21 phenylalanine 4.07 3.77 tryptophan 0.93 23.9 ______________________________________

5 more elements

A process for isolating protein PP.sub.4 which has

(a) an electrophoretic mobility in the range between that of 60 .sub.1 and .alpha..sub.2 globulins;
(b) an isoelectric point of 4.85.+-.0.15;
(c) a sedimentation coefficient s.sub.20,w.sup.O of 3.3.+-.2S;
(d) a molecular weight determined in polyacrylamide gel containing sodium dodecylsulfate (DSD) of 35,000.+-.5,000;
(e) an extinction coefficient E.sub.1 cm.sup.1% (280 nm) of 5.9.+-.0.6;
(f) a carbohydrate content of 2.4.+-.0.94% (g/100 g) (mannose 0.3.+-.0.2%, galactose 0.4.+-.0.2%, xylose 0.1.+-.0.04%, glucose 0.2.+-.0.1%, glucosamine 1.0.+-.0.2% and neuraminic acid 0.4.+-.0.2%) and
(g) the following aminoacid composition:
______________________________________ Residues per 100 residues Coefficient of Aminoacid (mol-%) variation ______________________________________ lysine 6.95 1.14 histidine 0.97 17.4 arginine 5.44 1.77 aspartic acid 11.41 1.68 threonine 6.78 2.40 serine 6.21 2.26 glutamic acid 12.25 0.43 proline 1.96 6.20 glycine 6.68 3.83 alanine 7.92 1.67 cystine 1/2 0.77 19.5 valine 5.34 3.80 methionine 1.98 6.00 isoleucine 5.21 2.23 leucine 11.50 0.45 tyrosine 3.55 4.21 phenylalanine 4.07 3.77 tryptophan 0.93 23.9 ______________________________________
which comprises subjecting an extract isolated with dilute salt or buffer solutions from organs which contain this protein to one or more of the following measures:
(a) precipitating the protein PP.sub.4 with ammonium sulfate in the pH range from 5 to 8 and at 40-70% saturation;
(b) precipitating the protein PP.sub.4 with a water-soluble acridine base at a pH between 4 and 9 and at a concentration of the base of 0.2-0.8 g/100 ml;
(c) preparative zone electrophoresis which entails the protein fraction between .alpha..sub.1 and .alpha..sub.2 globulins being isolated;
(d) gel filtration or ultrafiltration which entails proteins in the molecular weight range of 20,000 to 50,000 being isolated;
(e) adsorption on kaolin and elution of the protein PP.sub.4 ;
(f) adsorption on a weakly basic ion exchanger and elution of the protein PP.sub.4 ;
(g) concentration by immunoadsorption.

13 more elements

Dependencies are inferred from the claim text.

Tissue protein PP.sub.4, a process for isolating it and its use

Life of this patent

Tissue protein PP.sub.4, a process for isolating it and its use

Life of this patent

Abstract

Claims4