Immune syntony scattering spectrometry for detecting B factor

Abstract

The invention discloses a resonance scattering spectrometric for detecting B factor, which utilizes the resonant scattering effect of the immune complex to detect. When the B factor in the blood serum is compounded with the specificity goat to human B factor antibody, an insoluble complex is formed, the insoluble complex has the resonant scattering effect, the insoluble complex congregates with the effect of the PEG, and the scattering intensity is strengthened obviously. The content of the B factor can be worked out by the B factor concentration of the calibration serum--resonance scattering correct value curve. Compared with the existing method, the invention combines the immune-reaction with the resonance scattering spectrometric technique; the invention has the advantages that the sensitivity is high and the detection limit is low, and a Mu g / ml level can be achieved, the dosage of the sample used for testing low concentration B factor is little, the operation is simple and convenient, the time of test is short, the reagent used for the test is innocuous and safe, and the cost is low.

Description

Technical field: [0001] The invention relates to a detection method for B factor, in particular to an immunoresonance scattering spectrometry for detecting B factor. Background technique: [0002] B Factor (BF), also known as C 3 Activator precursor (C 3 PA), which is an important component in the alternative complement pathway activation pathway. Beta 2 Globulin, with a molecular weight of 100,000 D, has only one polypeptide chain, is rich in glycine, and is unstable to heat. It will be inactivated within 30 minutes at 50°C. Under the action of D, factor B is cleaved into two fragments a and b with different antigenicity. B factor must be with C 3b Etc and Mg 2+ Combines into C in the presence of 3b·B After that, B is released by the action of D a , forming C 3b ·B b , the complex has C 3 Invertase activity, acting on C 3 generate more C 3b , forming under the condition of P factor (C 3b )nB b P, which is the C in the alternative pathway 5 Invertase. [000...

AI Technical Summary

Problems solved by technology

Among them, the classic immunoprecipitation method (one-way diffusion, rocket electrophoresis, etc.) can detect the content of factor B, but the one-way diffusion method has low sensitivity, poor accuracy and reproducibility, and the process is too long; the rocket electrophoresis method is complicated to operate The hemolysis method is often used to determine the activity of factor B; the double-antibody sandwich ELISA method avoids the labeling of specific antibodies, but increases the operation steps, and needs to use the enzyme-labeled antibody against the second animal, the cost of the reagent is high, and the used The enzyme substrates are often aniline and phenol reducing dyes, which are less stable and easily oxidized when exposed to light or air. What’s more, aniline substrates have carcinogenic effects, and the determination process includes coating, incubation, etc. , Washing, immune reaction, color development and other steps take a long time, at least 24 hours, and the operation is cumbersome

[0006] In recent years, some people have used immunoturbidimetry to measure factor B. Although it is simple and fast, the sensitivity is not high.

In addition, some researchers use the surface plasmon resonance B factor sensor to measure the B factor, although the detection limit is low, but the operation is complicated; there is also a piezoelectric immunosensor to measure the B factor, which has high sensitivity, selectivity and reproducibility. Both are good, but the operation process is long and complicated

The immunoresonance scattering spectrometry method of the present invention detects B factor and has not been reported yet

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