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Antibody combination for replacing lateral scattering light signal in mass spectrum flow type blood tumor immunotyping and application thereof

A technology for hematological tumor and immunophenotyping, applied in the field of mass flow cytometry, which can solve the problems of limited mass flow cytometry and inability to distinguish cell subsets.

Active Publication Date: 2022-07-08
浙江普罗亭健康科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Since mass spectrometry uses mass spectrometry methodology, the cells are completely ionized, so the traditional flow SSC is not included in the detection process, and when it is applied to hematological tumors, the first-level gate similar to flow cytometry cannot be performed. That is, the combined gate of CD45 and SSC cannot distinguish major cell subgroups, which limits the clinical application and scientific research of mass spectrometry in the field of hematological tumors. It is necessary to develop an antibody combination that can replace traditional SSC and can well Make up for this deficiency of mass spectrometry flow cytometry, and better utilize its advantages of multi-parameter simultaneous detection, so that mass spectrometry flow cytometry can be better applied to the immunophenotypic detection of hematological tumors

Method used

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  • Antibody combination for replacing lateral scattering light signal in mass spectrum flow type blood tumor immunotyping and application thereof
  • Antibody combination for replacing lateral scattering light signal in mass spectrum flow type blood tumor immunotyping and application thereof
  • Antibody combination for replacing lateral scattering light signal in mass spectrum flow type blood tumor immunotyping and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1 Immune typing of healthy human bone marrow cells

[0051] 1), prepare fresh healthy human bone marrow and remove mature red blood cells.

[0052] 2), take 1-3x10^6 cells, resuspend with PBS, adjust the volume to 1mL, add 50ul-1mL 194Pt (0.1-1μM), and stain at room temperature for 2min to distinguish between dead and live cells.

[0053] 3), add 2mL bovine serum albumin solution (including 375-625 mass parts of bovine serum albumin, 15-25 mass parts of sodium azide, 75-125 volume parts of phosphate buffer solution), centrifuge at 500g / 5min, remove by suction The supernatant was added with 50uL of blocking solution and blocked on ice for 20min, wherein the blocking solution consisted of 0.5μL of human immunoglobulin solution (including 15-25 parts by mass of human immunoglobulin, 0.15-0.25 parts by mass of sodium azide, 0.75-1.25 phosphate buffer solution), 0.5uL mouse immunoglobulin solution (including 15-25 parts by mass mouse immunoglobulin, 0.15-0.25 parts ...

Embodiment 2

[0063] Example 2 Immune typing of bone marrow cells in patients with acute lymphoblastic leukemia

[0064] 1), prepare the bone marrow of fresh acute lymphoblastic leukemia patients and remove mature red blood cells.

[0065] 2) Take 1-3×10^6 cells, resuspend with PBS, adjust the volume to 1mL, add 50ul-1mL 194Pt (0.1-1μM), and stain at room temperature for 2min to distinguish between dead and live cells.

[0066]3), add 2mL bovine serum albumin solution (including 375-625 mass parts of bovine serum albumin, 15-25 mass parts of sodium azide, 75-125 volume parts of phosphate buffer solution), centrifuge at 500g / 5min, remove by suction The supernatant was added with 50uL of blocking solution and blocked on ice for 20min, wherein the blocking solution consisted of 0.5uL of human immunoglobulin solution (including 15-25 parts by mass of human immunoglobulin, 0.15-0.25 parts by mass of sodium azide, 0.75-1.25 phosphate buffer solution), 0.5uL mouse immunoglobulin solution (includi...

Embodiment 3

[0076] Example 3 Immune typing of bone marrow cells in patients with acute myeloid leukemia

[0077] 1) Prepare the bone marrow of fresh acute myeloid leukemia patients and remove mature red blood cells.

[0078] 2), take 1-3x10^6 cells, resuspend with PBS, adjust the volume to 1mL, add 50ul-1mL 194Pt (0.1-1μM), and stain at room temperature for 2min to distinguish between dead and live cells.

[0079] 3), add 2mL bovine serum albumin solution (including 375-625 mass parts of bovine serum albumin, 15-25 mass parts of sodium azide, 75-125 volume parts of phosphate buffer solution), centrifuge at 500g / 5min, remove by suction The supernatant was added with 50uL of blocking solution and blocked on ice for 20min, wherein the blocking solution consisted of 0.5uL of human immunoglobulin solution (including 15-25 parts by mass of human immunoglobulin, 0.15-0.25 parts by mass of sodium azide, 0.75-1.25 phosphate buffer solution), 0.5uL mouse immunoglobulin solution (including 15-25 pa...

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Abstract

The invention discloses an antibody combination for replacing a lateral scattering light signal in mass spectrum flow type blood tumor immunotyping. The antibody combination comprises a Lactoferrin antibody and a Lysozyme antibody. The invention also discloses a portal method for mass spectrum flow type blood tumor immune typing. The invention further discloses a kit for mass spectrum flow type blood tumor immune typing. According to the invention, a Lactoferrin antibody and a Lysozyme antibody are adopted for the first time, a two-stage enclosure strategy is carried out in combination with a CD45 antibody, and a mass spectrometry flow cytometry is matched, so that the traditional flow CD45 / SSC can be replaced to distinguish mature granulocytes, mononuclear cells, nucleated red blood cells, lymphocytes, primitive and immature cells, abnormal cell subgroups and other subgroups in bone marrow; the technical problem that the SSC cannot be detected in the blood tumor cell analysis of a mass spectrum flow type is broken through, and the immunotyping depth of the current blood tumor can be improved by combining the multi-parameter high-throughput characteristic of the mass spectrum flow type.

Description

technical field [0001] The invention relates to the technical field of mass flow cytometry, in particular to an antibody combination and application for replacing side scattered light signals in the mass flow cytometry blood tumor immunophenotyping. Background technique [0002] Accurate typing of hematological tumors is the premise for the correct selection of treatment options. Currently, cell morphology, immunology, cytogenetics and molecular biology typing, that is, MICM typing, are commonly used internationally. Among them, multi-parameter flow cytometry for immunological typing plays an important role. This method improves the accuracy of identifying specific disease types based on the immune markers of patient tumor cells. [0003] Multiparameter flow cytometry detects CD molecules on the surface of bone marrow cells by fluorescent antibodies, such as stem / progenitor cell antigens, bone marrow cell line-related antigens, red blood cells, B cells, T cells, NK cells, me...

Claims

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Application Information

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IPC IPC(8): G01N15/14G01N27/62
CPCG01N15/14G01N27/62G01N2015/1006A61K2039/505C07K16/40C07K16/18G01N33/6848G01N33/56966G01N15/10G01N2015/1021G01N33/57407G01N33/57484G01N2015/012
Inventor 林铖仇陈盛
Owner 浙江普罗亭健康科技有限公司
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