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Reagent kit and method for detecting tumor recurrence and transfer and evaluating curative effect

A technology for tumor metastasis and kits, which can be used in measurement devices, instruments, scientific instruments, etc., and can solve problems such as protein sequence identification and analysis, difficult chemical structures, etc.

Inactive Publication Date: 2008-06-04
许洋
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, many substances in cells often exist in trace amounts. The current methods for analyzing proteins have limitations in the above aspects. It is difficult to identify and analyze chemical structures and protein sequences with these conventional methods.

Method used

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  • Reagent kit and method for detecting tumor recurrence and transfer and evaluating curative effect
  • Reagent kit and method for detecting tumor recurrence and transfer and evaluating curative effect
  • Reagent kit and method for detecting tumor recurrence and transfer and evaluating curative effect

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1 A kit and method for detecting tumor recurrence, metastasis and evaluating curative effect

[0054] (1) Experimental method

[0055] 1. Materials

[0056] 1. Specimen sources: blood samples from lung cancer, esophageal cancer, gastric cancer, liver cancer, colon cancer, breast cancer, cervical cancer, brain tumor, lymphoma, ovarian cancer, prostate cancer, pancreatic cancer, bladder cancer, nasopharyngeal cancer, etc. (serum and plasma). A. 50 tumors no transfer Patient's serum; B. 20 tumors transfer Patient's serum; C. 30 cases of different tumors relapse Patient's serum; D. 20 patients with recurrent and metastatic tumors After effective treatment serum.

[0057] 2. Quality control: A. Human standardized quality control serum B. Mass spectrometer laser energy regulation: Before each test, use the above-mentioned standardized quality control serum.

[0058] 2. Method

[0059]1. Collection of samples: After the whole blood is collected, draw the se...

Embodiment 2

[0072] Example 2 Molecular Identification of Serum Amyloid A in Blood

[0073] Carbodiimide method (Carbodiimide Method) will have carboxylic acid group labeled magnetic bead on the matrix and the amino group of anti-serum amyloid A antibody (Gunn DL, et al.Preparation of sensitive and stable erythrocytes by the carbodiimidc method for the detection of primary and secondary IgM and IgG antibody. J Immunol Methods. 1972; 1(4): 381-389.).

[0074] Samples were spotted on the site of an anti-serum amyloid A antibody matrix.

[0075] Wash with binding buffer. Apply the first wash solution to the site before the sample is completely dry. The wash solution was left on the spot for at least 10 seconds. Thoroughly remove the first wash solution and repeat the above steps with the second wash solution. Thoroughly wash the entire array point with 1% trifluoroacetic acid, elute the biomarkers onto a special metal sheet for mass spectrometry (with 3×3mm round holes), dry the metal she...

Embodiment 3

[0084] Example 3 Sorting identification of mutated serum amyloid A

[0085] The 11439±15, 11527±15 and 11683±15 Da biomarkers were sequenced using multiple-stage mass spectrometry (MS / MS), post-source fragmentation (PSD) and protein ladder sequencing. By breaking molecules into pieces, protein ladders can be generated. This gradient is then analyzed by mass spectrometry. The 11439±15, 11527±15 and 11683±15 Da biomarkers were identified as variant serum amyloid A. Its chemical structure is (104 amino acids arranged from N-terminal to C-terminal):

[0086] Chemical structures of 11683 ± 15 Da biomarkers:

[0087] N-terminal

[0088] RSFFSFLGEAFDGARDMWRAYSDMREANYIGSDKYFHARGNYDAAKRGPGGVWAAEAISDARENIQRFFGHGAEDSLADQAANEWGRSGKDPNHFRPAGLPEKY

[0089] C-terminal.

[0090] Chemical structure of the 11527±15 Da biomarker:

[0091] N-terminal

[0092] SFFSFLGEAFDGARDMWRAYSDMREANYIGSDKYFHARGNYDAAKRGPGGVWAAEAISDARENIQRFFGHGAEDSLADQAANEWGRSGKDPNHFRPAGLPEKY

[0093] C-terminal.

[0...

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Abstract

The invention relates to a kit and a novel method for detecting tumor metastasis and recurrence and evaluating healing effect, which is a detecting method of non-invasive critical protein in vitro. The method finds a novel variant serum amyloid A by identifying and detecting with an antibody to attach the surface matrix and the mass spectrometric method; the variant serum amyloid A or peak values of 11439+-15, 11527+-15 and 11683+-15Da can be used for detections of tumor metastasis and recurrence and the evaluation of healing effect. The higher variant serum amyloid A or peak values of 11439+-15, 11527+-15 and 11683+-15Da prompt that the tumor is transferred and recurred and the healing effect is poor. The method provided by the invention can detect different tumor metastases and recurrences and evaluate the healing effect with 100 percent sensitivity and 100 percent specificity. The invention receives the variant serum amyloid A through a group of peak values of 11439+-15, 11527+-15 and 11683+-15Da or the surface matrix attached by the antibody, detects with quantitative mass spectrum analysis under the control of standard quality control serum, and can be applied to the detection method or kit development of biomarker combination in fluid which is separated from human body. The method is accurate, convenient and quick.

Description

technical field [0001] The present invention relates to a new method for non-invasive detection and evaluation of kits for critically ill patients, which is characterized in that it uses mass spectrometry to accurately identify and detect serum amyloid A that has been captured by WCX / SAX magnetic beads or antibodies Methods. The method provided by the invention can detect tumor metastasis and recurrence, and evaluate curative effect of tumor patients, and the sensitivity is 100%, and the specificity is 100%. The mutated serum amyloid A is an early warning protein fingerprint group for early detection of tumor progression, prediction of metastasis and recurrence. Its continuous negative expression can indicate that the condition of the tumor patient is stable, and its turning from positive to negative after treatment can also achieve this purpose. Therefore, using it as a clinical detection platform to predict the development of tumors and suggest corresponding treatment has ...

Claims

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Application Information

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IPC IPC(8): G01N30/02G01N33/543G01N33/577
Inventor 许洋
Owner 许洋
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