Waterborne adhesive chromatography media and method of using waterborne adhesive chromatography media for detection

Abstract

The invention discloses a waterborne adhesive chromatography media. The waterborne adhesive chromatography media comprises polysucrose, gelatin and water, wherein the concentration of the polysucrose is 5-100g / L, and the concentration of the gelatin is 1-10g / L. The waterborne adhesive chromatography media can be used for separating red blood cells and free immune globulin so as to detect incomplete antibodies and separating particulate antigen-antibodies so as to detect antigen-antibodies. A method of using the waterborne adhesive chromatography media for detection can be applied to the detection of blood cells such as red blood cells, blood platelets and white blood cells and can also be used for separating other particulate antigens and free soluble protein in immunolabelling technologies such as ELISA (enzyme-linked immuno sorbent assay), immunofluorescence and flow cytometry assay, so that a complicated washing process is avoided in an operation process of an immunology method, operation steps are greatly simplified, and the detection efficiency is improved; the waterborne adhesive chromatography media can be used for the detection of lots of samples; raw materials of the waterborne adhesive chromatography media are low in cost; the waterborne adhesive chromatography media has low requirements on a production technology.

Description

technical field [0001] The invention relates to the field of medical detection, in particular to a water-based gel chromatography medium and a detection method. Background technique [0002] After the incomplete antibody (mainly IgG antibody) binds to the corresponding antigen of red blood cells (sensitized red blood cells), no visible agglutination reaction occurs in saline medium, but after adding anti-IgG antibody (secondary antibody), the secondary antibody Combined with the Fc segment of the incomplete antibody on the sensitized red blood cells, it can be bridged and an agglutination reaction occurs. This test is called anti-human globulin test or Coombs test. It is used in routine clinical antibody screening, identification and In cross-matching, incomplete antibody testing is required for both recipients and donors who have a history of blood transfusion, pregnancy, and transfusion of blood products. The detection of incomplete antibodies is one of the most important...

AI Technical Summary

Problems solved by technology

In this regard, the traditional test tube anti-human globulin test was invented in 1945, and it has been used up to now and can only be used in the confirmation test of a small number of specimens. Due to its cumbersome operation, it has never become a routine clinical detection method; currently the most widely used microcolumn gel Antihuman globulin test, which does not require a washing process, but has the disadvantages of high reagent prices and false positives for fibrinogen in blood samples; in addition, various "enhanced tests", including protease, albumin, polybrene etc., because of their respective shortcomings, they have not been able to become a reliable method for detecting blood group incomplete antibodies

[0003] In 1945, scientists such as Coombs RRA believed that their blood should contain anti-erythrocyte agglutinin based on the symptoms of some patients. However, no agglutination reaction occurred after mixing these serums with red blood cells. ), the agglutination reaction occurs when it is used as a bridge. Initially, this method was only used to detect incomplete antibodies in serum. After development, it is also used to detect red blood cells that have combined antibodies and / or complements in patients, that is, the classic Coombs test. The former It is an indirect anti-human globulin test, and the latter is a direct anti-human globulin test. The classic Coombs test should be a deterministic and standard test for incomplete antibody detection now and in the future. Routine incomplete antibody detection and detection of small molecule antigen antibodies other than red blood cells, but because anti-human spheres can combine with antibodies to sensitized red blood cells in serum and free immunoglobulins in serum, so in the conventional test tube Coombs test First, wash to remove free immunoglobulins that are not combined with red blood cells, and then add anti-human spheres, so as to avoid false negative reactions caused by the neutralization of anti-human spheres by non-specific immunoglobulins in the reaction solution, which requires centrifugal washing with normal saline At least three times, so that the non-specific immunoglobulin residue in the reaction solution is less than 1 / 5000 of the original amount, which leads to cumbersome and time-consuming operations, and some weak reaction antibodies cannot be detected, which cannot meet the requirements of clinical blood group matching. Due to the time requirement of the capacitive test, this method has not been able to become a routine clinical test item.

To solve this problem, some enhancing media such as polybrene, polyethylene glycol (PEG), low ionic strength saline (LISS) and enzyme solution have been developed. The media technology can reduce or avoid the washing procedure, reduce the incubation time and enhance the antigen-antibody reaction, but there are also some disadvantages: for example, the polybrene method is easy to cause false negatives, and the operation process is not easy to standardize; Anti-complement components in human globulin reagents can cause false-positive reactions; some antibodies (anti-K) bind to red blood cells much less in low-ionic solutions than in normal saline, so test in low-ionic solutions Sometimes it will lead to the missed detection of the antibody; enzyme treatment of red blood cells may cause false negative reactions due to the destruction of some blood group antigens, and it may also be due to the digestion of red blood cell membrane polypeptide molecules by proteases, making them expose hidden antigens, and then interact with anti-human False positive reaction of heterologous antibodies in globulin reagents, or polyagglutination antibodies in serum reaction solutions

Micro-column gel anti-human globulin test and solid-phase erythrocyte adsorption test can become experimental techniques for routine clinical application, but it is still impossible to completely replace the test tube Coombs test.

At present, there is no method that can detect all blood group antibodies 100%. Some antibodies are more complex, such as enzyme-only antibodies. Therefore, when encountering complex specimens, multiple methods should be used to coordinate processing to improve the detection of incomplete antibodies. Yield rate, to ensure clinical blood safety

[0005] In the development of antibody detection technology, from agglutination test to today's various immunolabeling techniques, such as ELISA for detecting particulate antigens, immunofluorescence, flow cytometry, etc., they have never been separated from the washing process, and the reaction process involves 2-3 Each reaction procedure needs to be washed more than 3 times, the operation is cumbersome and time-consuming, and the clinical application is limited

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