Sample preparation method for analysis of platelet proteins
View Patent ↗The present invention provides a novel method of in vitro sample preparation for the analysis of proteins. In a first aspect, the present invention is a method comprising the separation and individual treatment of blood platelets and platelet-poor plasma obtained from a platelet-rich plasma sample. This allows for optimal treatment of both fractions and overcomes many of the problems associated with current methods. In a second aspect of the present invention the platelet sample preparation method can be implemented before a biochip based immunoassay for the detection of platelet proteins in combination with plasma proteins.
1. A method of preparing an in vitro sample for protein analysis comprising the following steps:
(i) Centrifuging a platelet suspension and separating the resultant supernatant and pelleted platelets;
(ii) Separately treating the supernatant and pelleted platelets from step (i) with a lysis buffer or detergent; and
(iii) Combining the resulting lysates of step (ii)
wherein the platelet suspension is platelet-rich plasma.
2. The method of claim 1 wherein the supernatant is platelet-poor plasma.
3. The method of claim 1 wherein steps (ii) and (iii) include an incubation step, after the addition of lysis buffer or detergent in step (ii) and after combination of the lysates in step (iii).
4. The method of claim 1 , wherein the lysis buffer or detergent of step (ii) used to treat the pellet fraction is Sodium dodecyl sulphate buffer.
5. The method of claim 1 , wherein the protein analysis consists of an Immunoassay, protein array or 2D gel electrophoresis.
6. The method of claim 5 wherein said immunoassay is a biochip based immunoassay.
7. The method of claim 1 , wherein the lysis buffer is a radioimmunoprecipitation assay buffer (RIPA).