Heterogenous conjugate and application thereof to RV (rubella virus) assay

A technology of heterologous cross-linking and cross-linking, which is applied in the field of immunology, can solve problems such as difficult standardization, difficult inactivation, and unstable quality, and achieve the effects of reducing detection errors, simple production and preparation, and avoiding infectivity

Active Publication Date: 2015-05-27
GENCLONN BIOTECH HANGZHOU
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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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  • Heterogenous conjugate and application thereof to RV (rubella virus) assay
  • Heterogenous conjugate and application thereof to RV (rubella virus) assay
  • Heterogenous conjugate and application thereof to RV (rubella virus) assay

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 heterogenous conjugate. The heterogenous conjugate comprises a tetramer shaped heterogenous crosslinking antibody formed by mouse anti-RV (rubella virus) McAb and healthy human IgG (immunoglobulin G) which is not infected with RV, wherein the antibody can serve as a substitute of an RV antibody in the positive blood of patients and can be used in immunoassay kits as a quality control product replacing the positive blood of patients after being diluted in a diluent. The heterogenous conjugate is simple to produce and prepare, is low in cost, has very good safety and stability at the same time and can conduce to reducing the assay errors.

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 The application of the rubella virus antibody substitute in the 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-resolve...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/46G01N33/569
Inventor 潘剑用周海涛闵丹
Owner GENCLONN BIOTECH HANGZHOU
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