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Method for assessing cell surface receptors of blood cells

A cell surface, blood cell technology applied to one or more subtypes of blood cells that can solve problems such as operator error

Inactive Publication Date: 2018-04-17
GENTIAN DIAGNOSTICS AS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, it is a 40-minute test that also requires manual addition of a separate liquid reagent after 17 minutes, which is prone to operator-induced error during the test
In addition, built-in magnetic separation devices and complex production of magnetic particles for the process separation steps are required

Method used

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  • Method for assessing cell surface receptors of blood cells
  • Method for assessing cell surface receptors of blood cells
  • Method for assessing cell surface receptors of blood cells

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Experimental program
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Embodiment approach

[0101] B1. The present invention relates to the following specific embodiments:

[0102] 1. An assay method for assessing one or more subclasses of blood cells of interest (BCoI) in a liquid whole blood sample or a sample derived therefrom, each subclass carrying a first, preferably distinguishable, target blood cell cell surface markers (or cell surface receptor molecules) (M1) of said subclass of said subclass, which means that the markers (M1) for different cell subclasses are different from each other (i.e. antigenically different and thus distinguishable),

[0103] wherein said sample may additionally comprise (or be suspected of comprising) interfering blood cells (DBC) carrying at least one of said first cell surface markers (M1) as non-specific markers and thus interfering also carrying at least one of said markers (M1) Assessment of said subclass of BCoI, and / or wherein said sample may additionally contain (or be suspected of containing) at least one free (e.g. lysed)...

Embodiment 1

[0329] Example 1: Preparation of nitrocellulose filters with an average pore size of 3, 5, and 8 μm and nylon mesh filters with an average pore size of 30 μm

[0330] Whatman nitrocellulose filters (Cat. No. 7193-002 for 3 μm pore size, Cat. No. 7195-004 for 5 μm pore size, and Cat. No. 10400112 for 8 μm pore size) and Millipore nylon mesh filters (Cat. Soak in bovine serum albumin aqueous solution for 4 hours. This blocking process is performed to avoid non-specific binding of proteins and cells to the filter when later used in a vertical filter device.

[0331] Preferably, since the nylon mesh filter is a fluffy material, the nylon mesh filter can be supported in the periphery by a ring of polystyrene or another harder material. The stiffer material should be glued (e.g., by Clearsol Casco glue) or welded to the nylon mesh filter to prevent liquid from leaking between the ring and the nylon mesh filter (see below in Example 7. figure 1 filter (106) and ring (108) in).

Embodiment 2

[0332] Example 2: Preparation of Polyclonal Antibody Anti-human CD14 Receptor Antibody from Chicken Eggs

[0333] Human CD14 receptor, customized by Novoprotein Inc, US, has the following amino acid sequence (SEQ ID NO: 1)

[0334]

[0335] It was suspended in Freund's complete adjuvant (FCA).

[0336] Polyclonal anti-human CD14 antibodies can be prepared by methods known in the art, for example, Chase, M.W., 1967 in "Methods of Immunology and Immunochemistry", Williams, A. et al., M.W., pp. 197-209, Academic Press, New York those described in . Briefly, an animal of a suitable species (for example rabbit, goat or sheep, or preferably avian, especially poultry, such as hen). Following immunization, animals are bled and polyclonal antibodies are purified by methods such as ammonium sulfate or ammonium chloride precipitation, anion exchange chromatography, immunoaffinity chromatography, and / or affinity chromatography.

[0337] Polyclonal avian antibodies are usually obt...

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Abstract

The present invention relates to a novel method for rapid assessment of one or more subclasses of blood cells of interest (BCol), as for example CD4+ cells and CD8+ cells, in a liquid whole blood sample or a sample derived therefrom; a method of determining the cell count for such cells; a method for determining the CD4 / CD8 ratio; a method for determining the quantity of such receptors in a sample; as well as a vertical flow assay device for performing such assessment.

Description

[0001] The present invention relates to: novel methods for the rapid assessment of one or more subclasses of blood cells of interest (BCoI), such as CD4+ cells and CD8+ cells, in liquid whole blood samples or samples derived therefrom; A method of determining the cell count of the cells; a method of determining the CD4 / CD8 ratio; a method of determining the amount of the receptor in a sample; and a vertical flow assay device for making this assessment. Background technique [0002] Whole blood is the term used for human blood from a standard blood donation or blood sampling. During collection, blood is usually combined with an anticoagulant, but is generally not otherwise processed. Whole blood contains plasma, red blood cells (erythrocytes) and white blood cells (leukocytes), and platelets. Heparin, citrate, and EDTA (ethylenediaminetetraacetic acid) are common anticoagulants added to prevent clotting in blood samples for laboratory analysis. [0003] CD4+ T helper cells ar...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/569
CPCG01N33/56972G01N33/54391G01N33/58G01N33/6872G01N2333/70514G01N2333/70517
Inventor 埃尔林·松德列哈根
Owner GENTIAN DIAGNOSTICS AS
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