Method for detection of biological activity of Fc region of human immune globulin through ultraviolet spectrophotometry

Abstract

The invention discloses a method for detection of biological activity of the Fc region of human immune globulin through ultraviolet spectrophotometry. According to the method, an adsorbed diphtheria-acellular pertussis-tetanus vaccine is used to substitute high-cost hardly-available labelled antigen for sensitization erythrocytes; an Ultrospec 6300pro ultraviolet/visible light spectrophotometer produced by GE Company is used for detection of a sample, and software of the spectrophotometer carries out data processing. Experimental results of the invention prove that standard deviation is in a range of 0.06 to 15.92% and variance is in a range of 0.00 to 1.69%. Results of comparison between the results of an ultraviolet spectrophotometer and the mean value of three detection results of the method provided by the invention are as follows: average data deviation is -1.37%, standard deviation of data difference values is -1.28%, and variance of data difference values is 0.02%. The detection results of the invention is in a reasonable theoretical scope, so the method provided by the invention is indeed a cheap method for micro-scale detection of biological activity of the Fc region of human immune globulin in a blood product.

Description

technical field [0001] The invention belongs to the technical field of medicine, and relates to a method for detecting the biological activity of the Fc segment of human immunoglobulin at low cost and in a small amount in blood products, that is, a method for detecting the biological activity of the Fc segment of human immunoglobulin by ultraviolet spectrophotometry. Background technique [0002] Human Immunoglobulin G (IgG), an antibody molecule, accounts for 75% of plasma immunoglobulins and is synthesized and secreted by plasma B cells. Immunoglobulin Fc fragment (Fragment crystallizable region), which is the part of the antibody tail region that interacts with cell surface receptors and certain complement system proteins, can activate the immune system and is related to the effectiveness of IgG drugs. Purification during production, virus inactivation, ultrafiltration, filtration, and product storage can all lead to reduced biological activity and degradation of IgG mole...

AI Technical Summary

Problems solved by technology

From 2008 to 2011, the wholesale supply of various IgG blood products in my country was more than 51 million bottles, but the measurement of the biological activity of the Fc segment of IgG has not yet been carried out

Current IgG Fc biological activity determination, for example: "European Pharmacopoeia 7.0" uses rubella antigen and tanned packed red blood cells to prepare sensitized red blood cells for the detection of IgG Fc biological activity, this method requires rubella antigen coagulation unit requirements More than 256 units, and the concentrated rubella antigen is difficult to buy domestically. The price of foreign product Aalto Bio AW6088 rubella virus antigen 1mg is about 15,000 RMB, and its detection cost is very alarming; The packed erythrocytes tanned by virus are used for the detection of IgG biological activity. The disadvantage of this method is that there is no way to increase the detection throughput and only a single sample can be detected, which is a waste of manpower and material resources, which is not conducive to the popularization of IgG activity detection. The second disadvantage is diphtheria There are problems with the source of raw material reagents in the use and purchase of toxins, and the use of mumps virus requires a safety level BSL-2 biosafety laboratory, which requires high professional technical operations and laboratory hardware conditions, and is not conducive to the biological activity of the Fc segment of IgG. Popularization of detection; In 2000, RamasamyI et al. discovered the flow cytometry method to evaluate the functional activity of the Fc fragment of IgG, which requires high professional technical operation and laboratory hardware conditions, and is not conducive to the popularization of the biological activity detection of the Fc fragment of IgG; In 2004, Vrdoljak A et al. used tetanus toxoid as an antigen to replace the high-titer rubella virus antigen, which was difficult to obtain, to determine the biological function of the Fc segment of IgG preparations; in 2009, Georgakopoulos T et al. used frozen red blood cells instead, requiring continuous fresh A method for measuring the biological function of the Fc segment of IgG preparations on human red blood cells; in 2012, Georgakopoulos T et al. used Kodocyte technology to coat CMV antigens on red blood cells to obtain standardized preparations of sensitized red blood cells for rapid detection of the Fc segment of IgG preparations

[0006] The above methods have been explored to varying degrees in the detection of viral antigen labeling of sensitized red blood cells and the detection throughput of the Fc segment of IgG, but they have failed to establish a cheap and high-throughput detection of the Fc segment of IgG. method of biological activity

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