Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving virus or bacteriophage
Patent
1995-08-10
1997-07-22
Knode, Marian C.
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving virus or bacteriophage
435 71, 435 792, 436518, 436531, 530350, 530395, C12Q 170, G01N 3353, G01N 33543, C07K 1400
Patent
active
056502699
DESCRIPTION:
BRIEF SUMMARY
This application is a 371 of PCT/FR94/00142 filed Feb. 9, 1994.
The present invention relates to a method for detecting and/or assaying viral compounds and to a support bearing at least one glycoprotein.
According to the present invention, the term viral compound refers both to the compounds, in particular the proteins, constituting a virus and to particles of viral type. Particles of viral type are either virions, which may be complete or incomplete, or portions of virion, or assemblies containing constituent compounds of viruses, which have certain properties of viruses or of viral compounds; in particular, they are detected by certain antibodies specific for viral compounds or may be similar to these viral compounds.
According to the present invention, it has been found that certain viral compounds bound themselves specifically to one glycoprotein: .beta.2'-glycoprotein I or a protein composition containing .beta.2'-glycoprotein I. .beta.2'-Glycoprotein I is the glycoprotein described in the French patent application filed under No. 93 01399 by the same applicants on 9 Feb. 1993. .beta.2'-Glycoprotein I binds to the membrane of viral particles, as well as to certain chemical or biological surfaces or compounds. According to the present invention, this property is used in order to uptake and isolate viral compounds, thereby allowing them to be detected and/or assayed.
The subject of the present invention is thus a method for detecting and/or assaying viral compounds, characterized in that the viral compounds are bound using .beta.2'-glycoprotein I.
According to the invention, .beta.2'-glycoprotein I may be used pure or in the form of a protein composition containing in particular other glycoproteins, possibly other .beta.2'-glycoproteins. This composition may, in particular, be that obtained by elution of an affinity column on a gel bearing sulfate groups as described in French patent application 93 01399.
According to a first embodiment of the method, .beta.2'-glycoprotein I is hound to a support, a viral compound contained in a biological fluid or extracted from a biological tissue is then bound to the .beta.2'-glycoprotein I so as to separate the said viral compound from the compounds which are not capable of binding to .beta.2'-glycoprotein I, and the viral compound bound to the .beta.2'-glycoprotein I is detected and/or assayed by any method for detecting and/or assaying this vital compound.
According to a second embodiment of the method, the viral compound is bound to a support and is exposed with .beta.2'-glycoprotein I conjugated with a label. The viral compound may be bound either directly to the support or indirectly, for example via an antibody. The label may advantageously be an enzyme or a radioactive product.
.beta.2'-Glycoprotein I is the glycoprotein isolated from the residue bound to the affinity chromatography column(s) used in the method for the purification of albumin from blood plasma, described in FR-A 2,690,444. According to this document, the albumin is separated from the other proteins by a liquid phase chromatographic method in which the aqueous solution containing the albumin is passed through at least one so-called "hydrophobic" chromatography column filled with a particulate material capable of retaining some of the proteins other than albumin; in order to complete the separation, it is also proposed to pass the aqueous albumin solution through at least one affinity chromatography column containing a neutral particulate support or a particulate support which is close to neutrality charged with a polysulfated compound. The effluent obtained by this method consists of a solution of purified albumin, the majority of the proteins other than the albumin being bound either to the hydrophobic chromatography column(s) or to the affinity chromatography column(s).
This method is carried out, for example, as follows:
A raw aqueous solution of albumin is preferably obtained from blood plasma, in particular human plasma, by a known process of COHN type, according to which the plasma is fr
Graafland Hubert
Rucheton Marcel
Stefas Elie
Auer Henry E.
Knode Marian C.
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