A quantitative analysis kit for Glypican-3 and its preparation method

Abstract

The invention discloses a glypican-3 quantitative analysis kit. The glypican-3 quantitative analysis kit comprises a GPC3 standard product, an acridinium ester-marked glypican-3 antibody solution, and a nano-magnetic microsphere solution which is coupled with glypican-3 antibodies. The invention further provides a preparation method for the kit. The GPC3 nano-magnetic microsphere chemiluminescence immunoassay determination kit which takes acridinium ester as a luminescence marker provided by the invention has the characteristics of high sensitivity, wide linear range, high specificity, stable marker, low cost and the like; the detecting process of the kit is simple and quick; the glypican-3 can be detected by only requiring a micro sample.

Description

technical field

[0001] The invention relates to a detection kit, in particular to a GPC3 detection kit. Background technique

[0002] Glypican-3 (glypican-3, GPC3) is a heparan sulfate-like proteoglycan, a complex carbohydrate complex covalently linked by proteins, lipids and sugars. GPC3 plays an important role in the formation and development of embryonic and fetal tissues and organs. It mainly plays a negative regulatory role, preventing excessive growth of tissues and organs and regulating body size as a whole; liver parenchymal cells are significantly expressed throughout the fetal period , is not expressed in the adult liver, but it can be reactivated and highly expressed in the early stage of liver cancer. In benign liver diseases (hepatitis, fatty liver), GPC3 is not expressed or expressed in low expression in liver cholangiocarcinoma, so soluble GPC3 in serum It can be used as a specific marker of primary liver cancer and play a role in the differential diagnosis o...

Examples

Embodiment 1

[0033] A quantitative analysis kit for Glypican-3, comprising the following:

[0034] (1) GPC3 standard product;

[0035] (2) Glypican-3 antibody solution labeled with acridinium ester;

[0036] (3) Nano-magnetic microsphere solution coupled with Glypican-3 antibody;

[0037] This test kit is made by following steps:

[0038] (1) Preparation of GPC3 standard substance: GPC3 is formulated into the GPC3 solution of step concentration with standard substance buffer solution, and described standard substance buffer solution comprises that mass ratio is 1%BSA, 0.5%NaCl, 0.05%TritonX-100 or Tween20, 0.05% ProClin 300 and the balance of phosphate buffer; the concentration of the phosphate buffer is 0.1mol / L.

[0039] (2) Preparation of acridinium ester-labeled Glypican-3 antibody solution:

[0040] a. Dissolve the GPC3 antibody in a phosphate buffer solution with a pH of 6.8 and a concentration of 0.2 mol / L to a final concentration of 2 mg / L.

[0041] b. Add acridinium ester to ...

Embodiment 2

[0047] A quantitative analysis kit for Glypican-3, comprising the following:

[0048] (1) GPC3 standard product;

[0049] (2) Glypican-3 antibody solution labeled with acridinium ester;

[0050] (3) Nano-magnetic microsphere solution coupled with Glypican-3 antibody;

[0051] This test kit is made by following steps:

[0052] (1) Preparation of GPC3 standard substance: GPC3 is formulated into the GPC3 solution of step concentration with standard substance buffer solution, and described standard substance buffer solution comprises that mass ratio is 5%BSA, 1.0%NaCl, 0.1%TritonX-100 or Tween20, 0.1% Proclin-300 and the balance of phosphate buffer; the concentration of the phosphate buffer is 0.2mol / L.

[0053] (2) Preparation of acridinium ester-labeled Glypican-3 antibody solution:

[0054] a. Dissolve the GPC3 antibody in a phosphate buffer solution with a pH of 7.3 and a concentration of 0.2 mol / L to a final concentration of 8 mg / L.

[0055] b. Add acridinium ester to th...

Embodiment 3

[0061] A quantitative analysis kit for Glypican-3, comprising the following:

[0062] (1) GPC3 standard product;

[0063] (2) Glypican-3 antibody solution labeled with acridinium ester;

[0064] (3) Nano-magnetic microsphere solution coupled with Glypican-3 antibody;

[0065] (4) Luminous liquid.

[0066] This test kit is made by following steps:

[0067] (1) Preparation of GPC3 standard substance: GPC3 was formulated into 0ng / mL, 0.5ng / mL, 2.0ng / mL, 5.0ng / mL, 20.0ng / mL, 100.0ng / mL serial concentrations with standard buffer solution; Described standard buffer solution comprises mass ratio and is 3%BSA, 0.6%NaCl, 0.1%TritonX-100 or Tween20, 0.05%Proclin-300, the 0.2mol / L phosphate buffer of surplus, the pH of described standard buffer is 7.0.

[0068] (2) Preparation of acridinium ester-labeled Glypican-3 antibody solution:

[0069] ①. Dissolve the GPC3 antibody in phosphate buffer solution with a pH of 7.0 and a concentration of 0.2 mol / L to a final concentration of 5 mg...

technical field

[0001] The invention relates to a detection kit, in particular to a GPC3 detection kit. Background technique

[0002] Glypican-3 (glypican-3, GPC3) is a heparan sulfate-like proteoglycan, a complex carbohydrate complex covalently linked by proteins, lipids and sugars. GPC3 plays an important role in the formation and development of embryonic and fetal tissues and organs. It mainly plays a negative regulatory role, preventing excessive growth of tissues and organs and regulating body size as a whole; liver parenchymal cells are significantly expressed throughout the fetal period , is not expressed in the adult liver, but it can be reactivated and highly expressed in the early stage of liver cancer. In benign liver diseases (hepatitis, fatty liver), GPC3 is not expressed or expressed in low expression in liver cholangiocarcinoma, so soluble GPC3 in serum It can be used as a specific marker of primary liver cancer and play a role in the differential diagnosis o...

Examples

Embodiment 1

[0033] A quantitative analysis kit for Glypican-3, comprising the following:

[0034] (1) GPC3 standard product;

[0035] (2) Glypican-3 antibody solution labeled with acridinium ester;

[0036] (3) Nano-magnetic microsphere solution coupled with Glypican-3 antibody;

[0037] This test kit is made by following steps:

[0038] (1) Preparation of GPC3 standard substance: GPC3 is formulated into the GPC3 solution of step concentration with standard substance buffer solution, and described standard substance buffer solution comprises that mass ratio is 1%BSA, 0.5%NaCl, 0.05%TritonX-100 or Tween20, 0.05% ProClin 300 and the balance of phosphate buffer; the concentration of the phosphate buffer is 0.1mol / L.

[0039] (2) Preparation of acridinium ester-labeled Glypican-3 antibody solution:

[0040] a. Dissolve the GPC3 antibody in a phosphate buffer solution with a pH of 6.8 and a concentration of 0.2 mol / L to a final concentration of 2 mg / L.

[0041] b. Add acridinium ester to ...

Embodiment 2

[0047] A quantitative analysis kit for Glypican-3, comprising the following:

[0048] (1) GPC3 standard product;

[0049] (2) Glypican-3 antibody solution labeled with acridinium ester;

[0050] (3) Nano-magnetic microsphere solution coupled with Glypican-3 antibody;

[0051] This test kit is made by following steps:

[0052] (1) Preparation of GPC3 standard substance: GPC3 is formulated into the GPC3 solution of step concentration with standard substance buffer solution, and described standard substance buffer solution comprises that mass ratio is 5%BSA, 1.0%NaCl, 0.1%TritonX-100 or Tween20, 0.1% Proclin-300 and the balance of phosphate buffer; the concentration of the phosphate buffer is 0.2mol / L.

[0053] (2) Preparation of acridinium ester-labeled Glypican-3 antibody solution:

[0054] a. Dissolve the GPC3 antibody in a phosphate buffer solution with a pH of 7.3 and a concentration of 0.2 mol / L to a final concentration of 8 mg / L.

[0055] b. Add acridinium ester to th...

Embodiment 3

[0061] A quantitative analysis kit for Glypican-3, comprising the following:

[0062] (1) GPC3 standard product;

[0063] (2) Glypican-3 antibody solution labeled with acridinium ester;

[0064] (3) Nano-magnetic microsphere solution coupled with Glypican-3 antibody;

[0065] (4) Luminous liquid.

[0066] This test kit is made by following steps:

[0067] (1) Preparation of GPC3 standard substance: GPC3 was formulated into 0ng / mL, 0.5ng / mL, 2.0ng / mL, 5.0ng / mL, 20.0ng / mL, 100.0ng / mL serial concentrations with standard buffer solution; Described standard buffer solution comprises mass ratio and is 3%BSA, 0.6%NaCl, 0.1%TritonX-100 or Tween20, 0.05%Proclin-300, the 0.2mol / L phosphate buffer of surplus, the pH of described standard buffer is 7.0.

[0068] (2) Preparation of acridinium ester-labeled Glypican-3 antibody solution:

[0069] ①. Dissolve the GPC3 antibody in phosphate buffer solution with a pH of 7.0 and a concentration of 0.2 mol / L to a final concentration of 5 mg...

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