Methods for apolipoprotein detection
AssigneesBiocross, S.L., Instituto De Salud Carlos III, Centro De Investigacion Biomedica En Red De Enfermedades Neurodegenerativas
InventorsAndrés Rodríguez Martín, Miguel Calero Lara, Olga Calero Rueda, Luis García Albert
Life of this patent
Estimating the term…
Abstract
The present invention relates to methods for the detection and quantification of apolipoproteins and isoforms thereof in a sample, as well as to predictive methods of the probability of neurodegenerative or cardiovascular disease development based on apolipoprotein levels as determined by the detection methods of the invention.
Claims16
An in vitro method for detection and/or quantification in a sample of an apolipoprotein selected from the group consisting of apoAI, apoAIV, apoCI, apoCII, apoCIII, apoE and apoJ (clusterin) or an isoform thereof, the method comprising:
- (i) contacting the sample with a polystyrene surface under suitable conditions for a direct electrostatic and/or hydrophobic binding interaction between the apolipoprotein and the polystyrene surface, wherein the pH ranges from pH 2 to pH 10 during the contacting of the sample with the polystyrene surface;
- (ii) contacting the surface to which the apolipoprotein or the isoform thereof is bound in step (i) with an antibody specific for said apolipoprotein or for said isoform thereof under suitable conditions for formation of a complex between the antibody and the apolipoprotein or the isoform thereof;
- and
- (iii) detecting the complex formed in step (ii),
- wherein the polystyrene surface is blocked with albumin prior to contacting the sample.
An in vitro method for determining in a sample a relative amount of a target isoform of a given apolipoprotein selected from the group consisting of apoAI, apoAIV, apoCI, apoCII, apoCIII, apoE and apoJ (clusterin) with respect to total content of said apolipoprotein in the sample, the method comprising:
- (i) contacting the sample with a polystyrene surface under suitable conditions for forming a direct electrostatic and/or hydrophobic binding interaction between the apolipoprotein and the polystyrene surface;
- (ii) contacting the surface to which the apolipoprotein is bound as formed in step (i) with a first antibody specific for said apolipoprotein target isoform and with a second antibody which is capable of binding to all isoforms of said apolipoprotein that are present in the sample, wherein said contacting with said first and second antibodies is carried out under conditions suitable for formation of a first complex comprising the first antibody and the apolipoprotein target isoform and for formation of a second complex comprising the second antibody and all the isoforms of said apolipoprotein that are present in the sample;
- (iii) detecting a first level of the first complex formed in step (ii) and a second level of the second complex formed in step (ii);
- and
- (iv) determining relative amounts of said apolipoprotein target isoform with respect to the total apolipoprotein content based on the first and second levels detected in step (iii),
- wherein the polystyrene surface is blocked with albumin prior to contacting the sample.
Dependencies are inferred from the claim text.