Protein quantitation with cell imaging densitometry

Details Protein quantitation with cell imaging densitometry Protein quantitation with cell imaging densitometry

G - Physics

01

N

G01N 1/28 (2006.01) G01N 1/30 (2006.01) G01N 33/483 (2006.01) G01N 33/53 (2006.01) G01N 33/543 (2006.01) G01N 33/554 (2006.01) G01N 33/567 (2006.01) G01N 33/574 (2006.01) G02B 21/34 (2006.01) G01N 15/10 (2006.01) G01N 15/14 (2006.01)

Patent

CA 2347002

A method for quantitating cellular proteins in tissue, by means of a cell imaging densitometer in conjunction with immunohistological staining and a reference standard, is provided. Unlike prior art methods, which provide ordinal measures of relative amounts of protein among different cells, the method enables the quantitation of antigenic proteins in terms of absolute mass of protein/tumor or protein/patient, molecules of protein per cell, and volume or fraction of a tissue sample expressing the protein of interest. The method is useful for research purposes in the study of protein expression, and is shown to improve the accuracy of clinical histopathological analysis of tumor tissue sections for diagnosis and prognosis. Potentially useful for prescribing in situ treatment dosages. The demonstrated resulting improvement in the correlation between tissue levels and blood levels of tumor-associated proteins should facilitate minimally invasive monitoring of cancer progression and therapeutic response.

L'invention concerne un procédé permettant de quantifier les protéines cellulaires dans un tissu au moyen d'un densitomètre par imagerie cellulaire, d'une coloration immunohistologique et d'une norme de référence. Contrairement aux procédés déjà connus, qui proposent des mesures ordinales de quantités relatives de protéines parmi différentes cellules, ce procédé permet de quantifier des protéines antigènes en termes de masse absolue entre protéine/tumeur ou protéine/patient, de molécules de protéine par cellule et de volume ou fraction d'un prélèvement tissulaire exprimant la protéine d'intérêt. Ce procédé permet de faire des recherches dans le domaine de l'expression protéique et d'améliorer la précision des analyses histopathologiques cliniques de sections tissulaires tumorales en vue d'un diagnostic ou d'un pronostic. Ce procédé s'avère également utile pour prescrire des dosages de traitement in situ. L'amélioration apportée dans la corrélation entre la concentration tissulaire et la concentration sanguine de protéines liées à la tumeur devrait faciliter la surveillance avec effraction minimale de l'évolution du cancer et de la réponse thérapeutique.

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