Methods and reagents for improved selection of biological...

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G01N 33/553 (2006.01) C12N 5/00 (2006.01) C12Q 1/24 (2006.01) G01N 33/543 (2006.01)

Patent

CA 2493367

Methods, apparatus and compositions for separating a desired or undesired population or subpopulation from a biological sample are disclosed herein. The selection procedure is based on ferromagnetic, dense particles in a preferred size range from about 0.8 to about 1.2 microns. Specific binding agents are bound to the particles that recognize and bind to specific molecules on the targeted population or subpopulation, and the particles are mixed with the sample in such a way as to promote movement of the particles relative to the sample, promoting binding to the targeted population or subpopulation without non-specifically binding to non-targeted populations in the sample. Because of the large particle density, the bound population is separated from the fluid sample by gravity. Alternatively, the sample, including the bound, targeted population, is placed in a magnetic field such that the particles separate from the sample by evenly distributing over the vessel wall thus limiting non- specific trapping of the non~targeted population.

La présente invention a trait à des procédés, des appareils et des compositions permettant la séparation d'une population ou d'une sous-population souhaitée ou indésirable à partir d'un échantillon biologique. Le processus de sélection est basé sur des particules ferromagnétiques denses dans une plage de dimension préférée d'environ 0,8 à environ 1,2 microns. Des agents de liaison spécifique sont liés aux particules pour l'identification et la liaison à des molécules spécifiques sur la population ou sous-population ciblée, et les particules sont mélangées avec l'échantillon de manière à favoriser le déplacement des particules par rapport à l'échantillon, favorisant la liaison à la population ou sous-population ciblée sans liaison non spécifique aux populations non ciblées dans l'échantillon. Grâce à la grande densité des particules, la population liée est séparée de l'échantillon de fluide par gravité. En variante, l'échantillon comprenant la population ciblée liée est disposée dans un champ magnétique de sorte que les particules la séparent de l'échantillon par une répartition uniforme sur la paroi du réceptacle limitant ainsi le piégeage non spécifique de la population non ciblée.

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