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Human cytomegalovirus cross-linked antibody and its application in immunoassay kit

A technology of human cytomegalovirus and cross-linking agent, which is applied in the direction of hybrid immunoglobulin, measuring devices, instruments, etc., can solve problems such as difficult standardization, difficult inactivation, and unstable quality, so as to reduce detection errors and improve production and preparation. Simple, Avoid Infectious Effects

Active Publication Date: 2018-03-06
ACON BIOTECH (HANGZHOU) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are many disadvantages in using positive blood from patients to prepare positive quality control products: 1) It is difficult to obtain positive blood from patients with high titer and high specificity; ; 3) the cost is relatively high, 4) some pathogen infection cases are scarce, and it is difficult to obtain enough positive blood in a short period of time; At the same time, when collecting and inactivating the patient's positive blood, the operator also has the risk of potential infection
However, the above patents did not disclose the specific composition of the prepared heterologous cross-linked product, which caused the quality of the heterologous cross-linked product to be unstable between different batches, prone to detection errors, and poor substitution

Method used

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  • Human cytomegalovirus cross-linked antibody and its application in immunoassay kit
  • Human cytomegalovirus cross-linked antibody and its application in immunoassay kit
  • Human cytomegalovirus cross-linked antibody and its application in immunoassay kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] Utilize SPDP cross-linking agent (purchased from Thermo Scientific Company, product number 21857) to prepare the heterologous cross-linked product of mouse anti-HSV-II McAb and healthy human IgG not infected with HSV-II as HSV-II in patient's positive blood Antibody surrogate, the process is as follows:

[0045] (1) Dissolve 5 mg mouse anti-HSV-II McAb in cross-linking buffer (0.1M potassium phosphate, 0.1M NaCl, pH7.5), stir and add 50 μl SPDP cross-linking agent (3.2 mg / ml, dissolved in In absolute ethanol), after reacting at room temperature for 2 hours, put it into a dialysis bag, dialyze with reducing buffer (0.1M sodium acetate, 0.1M NaCl, pH4.5), and change the solution four times;

[0046] (2) Dissolve 5 mg of healthy human IgG uninfected with HSV-II in the cross-linking buffer, stir and add 50 μl SPDP cross-linking agent, react at room temperature for 2 hours, put it into a dialysis bag, and dialyze with the cross-linking buffer , change the liquid four times;...

Embodiment 2

[0053] Example 2 Experiments on substitution of mouse anti-HSV-II McAb and healthy non-HSV-II-infected human IgG heterologous cross-linked product, quality control stability and cryopreservation stability

[0054] Experiments were carried out using the mouse anti-HSV-II McAb prepared in Example 1 and the heterologous cross-linked product of healthy human IgG not infected with HSV-II as a substitute for the HSV-II antibody in the patient's positive blood.

[0055] (1) Preparation of quality control products. Prepare the heterologous cross-linked products in which the mass ratio of the tetrameric heterologous cross-linked antibody in the prepared heterologous cross-linked product is 50%, 70%, 90% and 100% in sequence as the patient Surrogates for HSV-II antibodies in positive blood. When the heterologous cross-linked antibody in the form of tetramer accounts for 50%, 70% or 90% of the mass ratio in the prepared heterologous cross-linked product, the remaining 50%, 30% or 10% he...

Embodiment 3

[0075] Example 3 Alternative experiment of mouse anti-HSV-1 McAb and healthy non-infected HSV-1 human IgG heterologous crosslinker

[0076] Using the SPDP cross-linker to prepare mouse anti-HSV-I McAb and healthy non-HSV-I-infected human IgG heterologous cross-linked product as a surrogate for HSV-I antibody in patient-positive blood, except for mouse anti-HSV -I McAb replaces mouse anti-HSV-II McAb, and the specific process is the same as in Example 1.

[0077] Experiments were carried out using the mouse anti-HSV-1 McAb prepared in Example 3 and the heterologous cross-linked product of healthy human IgG not infected with HSV-1 as a substitute for the HSV-1 antibody in the patient's positive blood. The heterologous cross-linked products in which the tetrameric heterologous cross-linked antibody accounts for 50%, 70%, 90% and 100% of the mass ratio in the prepared heterologous cross-linked product are sequentially prepared. When the heterologous cross-linked antibody in the f...

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Abstract

The invention provides a human cytomegalovirns cross-linking antibody. The human cytomegalovirns cross-linking antibody comprises a tetramer-form heterogenous cross-linking antibody formed by mouse anti-HCMV McAb and healthy human IgG which is not infected with HCMV. The invention further discloses an application of the antibody to preparing an HCMV immunoassay kit. The antibody provided by the invention is simple to produce and prepare, low in cost, and good in safety and stability. Besides, quality among quality control products of different batches is stable, so that the detecting errors can be effectively reduced.

Description

[0001] This application is a divisional application of the Chinese patent application with the application number 201310267573.6 and the application date on June 28, 2013. technical field [0002] The present invention relates to a heterologous cross-linked product, specifically a heterologous cross-linked product containing a heterologous cross-linked antibody in the form of a tetramer obtained from two heterologous antibodies through a cross-linking reaction, and Its application in the substitute of pathogen antibody in patient's positive blood belongs to the technical field of immunology. Background technique [0003] Pathogens such as viruses, bacteria, fungi and parasites pose an increasing threat to humans today. Immunoassay methods based on specific and highly sensitive reactions between antigens and antibodies, such as colloidal gold immunochromatography, latex immunochromatography, radioimmunoassay (RIA), enzyme-linked immunosorbent assay (ELISA), time-resolved Flu...

Claims

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Application Information

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IPC IPC(8): C07K16/46G01N33/569
Inventor 潘剑用周海涛闵丹
Owner ACON BIOTECH (HANGZHOU) CO LTD
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