Kit and application thereof in detection of vascular endothelial growth factor

Abstract

The invention discloses a kit and application thereof in detection of a vascular endothelial growth factor. The kit contains the following components which are independently stored: an antibody-coated magnetic bead working solution, a VEGF labeled antibody, a VEGF calibrator, a VEGF quality control product and a sample diluent. The antibody-coated magnetic bead working solution is a magnetic bead suspension coated with an antibody A. The VEGF labeled antibody is obtained by labeling an antibody B through an acridinium ester modifier. The acridinium ester modifier is acridinium sulfonyl hydrazide amine inner salt or other acridinium ester derivatives, and the antibody A and the antibody B are humanized VEGF antibody Fab segments with different amino acid sequences. The acridinium ester modifier is selected to be coupled with the humanized anti-VEGF antibody Fab segments, so that the stability is high, the sensitivity and the detection range of a detection system are improved, the storage time of the detection system is prolonged, and the reaction time is shortened. The linear range of the kit is 0.5-50000 pg / mL.

Description

technical field [0001] The invention belongs to the technical field of detection of vascular endothelial growth factor, in particular to a kit and its application in the detection of vascular endothelial growth factor. Background technique [0002] Vascular endothelial growth factor (VEGF) is a key regulator of physiological angiogenesis during embryogenesis, skeletal growth and reproductive function. Studies have found that VEGF is expressed in a variety of pathological and physiological conditions, so it can be used as a marker for in vitro diagnosis. At present, the in vitro detection kits for VEGF on the market mainly use quantum dot immunofluorescence method, enzyme-linked method, plate chemiluminescence method and magnetic particle chemiluminescence method. Existing test kit has the following problems at present: [0003] 1) The sensitivity and linear range need to be further improved. It has been reported that the VEGF content of healthy volunteers is 16.27±8.54pg / ...

AI Technical Summary

Problems solved by technology

If the sensitivity of the kit is not high, it cannot play a good role in early screening

At the same time, among the existing products, the product with the highest sensitivity is 0.1pg / ml, but the upper limit of its linear range is only 800pg / ml

However, the serum VEGF concentration range of IIIA / IIIB tumor patients is 14.9-1824.0pg / mL, which will lead to the dilution of high VEGF concentration samples in the actual detection, and the additional steps will reduce the reproducibility and reliability of the data. authenticity

However, the sensitivity of products with a relatively large detection linear range is insufficient (generally greater than 1pg / ml), which may also lead to a decline in the accuracy of detection data and affect clinical judgment

[0004] 2) The process is complicated

Both of these processes are relatively complicated, which is not conducive to the stable production of kits in large quantities.

[0005] 3) The detection time is longer

Compared with the two-step method, the total incubation time required by the one-step method is shorter, generally around 25 minutes, but the total incubation time required by the current one-step method is still relatively long, which cannot assist doctors to give disease control more quickly in the clinic plan

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