Kit for a Determination Method for Calprotectin and the Use of Calprotectin as a Predictive Marker for Cardiovascular Disease

Abstract

An assay kit for the use in a method for the detection of potential for CVD or propensity to CVD in a human or non-human animal subject. The method including assessing the concentration of calprotectin in a calprotectin-containing sample taken from the subject.

1 - 27 . (canceled) 
     
     
         28 . A kit for use in a diagnostic assay method comprising:
 one or more anti-calprotectin antibody- or antibody fragment-coated nanoparticles having a diameter in the range 65-140 nm,   wherein said assay method is an assay method for the determination of calprotectin in a calprotectin-containing body fluid, said method comprising the steps of;   
       (a) obtaining a calprotectin-containing liquid sample of, or derived from, said fluid; 
       (b) contacting said sample of said body fluid with said nanoparticle-bound anti-calprotectin antibody or antibody fragment, to bind said calprotectin; and 
       (c) assessing the calprotectin content by turbidimetry. 
     
     
         29 . The kit as claimed in  claim 28  further comprising a calprotectin solution of known concentration or a set of such solutions having a range of calprotectin concentrations. 
     
     
         30 . The kit as claimed in  claim 28  further comprising a light transmitting vessel. 
     
     
         31 . The kit as claimed in  claim 28  further comprising an opacification enhancer. 
     
     
         32 . The kit as claimed in  claim 28  further comprising a detector. 
     
     
         33 . The kit as claimed in  claim 28 , wherein the assay method is a direct turbidimetric assay method. 
     
     
         34 . The kit as claimed in  claim 28 , wherein the assay method is a direct turbidimetric assay method for the determination of calprotectin at concentrations of 0.5 to 50 mg/L in a calprotectin-containing body fluid. 
     
     
         35 . The kit as claimed in  claim 28  wherein in step a) said liquid sample comprises calprotectin at a concentration of 0.5 to 50 mg/L. 
     
     
         36 . The kit as claimed in  claim 28  wherein in said assay method increasing opacity correlates with increasing calprotectin concentration. 
     
     
         37 . The kit as claimed in  claim 28  wherein said assay is capable of measuring calprotectin with precision in samples having calprotectin concentrations throughout the range of 0.5-50 mg/L. 
     
     
         38 . The kit as claimed in  claim 28 , wherein said anti-calprotectin antibodies or antibody fragments are polyclonal having a population of calprotectin binding sites and are immobilised by binding or coupling, either directly or indirectly, to nanoparticles to form antibody or antibody fragment coated nanoparticles. 
     
     
         39 . The kit as claimed in  claim 37 , wherein said anti-calprotectin antibodies or antibody fragments are egg polyclonal antibodies or fragments thereof. 
     
     
         40 . The kit as claimed in  claim 28 , wherein said assay method is a direct turbidimetric assay method for the determination of calprotectin at concentrations of 0.5 to 50 mg/L in a calprotectin-containing body fluid, said method comprising the steps of:
 (a) obtaining a calprotectin-containing liquid sample of, or derived from, said body fluid, wherein said liquid sample comprises calprotectin at a concentration of 0.5 to 50 mg/L;   (b) contacting said liquid sample of said body fluid with anti-calprotectin antibodies or antibody fragments, to bind said calprotectin, wherein said anti-calprotectin antibodies or antibody fragments are egg polyclonal antibodies or fragments thereof having a population of calprotectin binding sites and are immobilised by binding or coupling, either directly or indirectly, to nanoparticles to form antibody or antibody fragment coated nanoparticles; and   (c) assessing the calprotectin content in said contacted liquid sample by turbidimetry,   wherein the diameter of the antibody or antibody fragment coated nanoparticles is 65-140 nm, wherein increasing opacity correlates with increasing calprotectin concentration; and wherein said assay is capable of measuring calprotectin with precision in samples having calprotectin concentrations throughout the range of 0.5-50 mg/L.   
     
     
         41 . The kit as claimed in  claim 28 , comprising two reagent mixtures only.