Reagent kit for inspecting apoAI and apoB

Abstract

In the invention, its dual dose types comprises: a) a reactant comprising buffer solution, high molecule accelerator; b) an antibody agent comprising anti apoA I antiserum, anti apoB antiserum; and antiserum buffer solution; c) fluid serotype constant calibration liquid comprising apoA I antigen, apoB antigen. Its single dose type comprises: a) antibody liquid comprising anti apoA I antiserum, anti apoB antiserum and reactant; b) liquid serotype constant calibrate liquid comprising buffer liquid, apoA I antigen and AapoB antigen.

Description

technical field [0001] The invention relates to a preparation method of an antibody and a kit for measuring serum components, in particular to a preparation method for anti-APOA I and APOB antibodies and a kit for measuring apoA I and apoB in serum, which can be widely used in medical and biochemical technologies field. Background technique [0002] Apolipoprotein A I (apo A I) is the largest component of the apoA family. ApoA I is a single polypeptide chain consisting of 243 amino acid residues with a molecular weight of 28300D. It is the main apolipoprotein of high-density lipoprotein (HDL) . HDL has the effect of preventing atherosclerosis, therefore, measuring the content of apoA I in serum (plasma) has become a routine detection item for preventing cardiovascular and cerebrovascular diseases. Apolipoprotein B (apoB) is the main protein of low-density lipoprotein (LDL), and increased LDL is a recognized risk factor for atherosclerosis. Therefore, the determination of s...

AI Technical Summary

Problems solved by technology

[0003] Known methods for measuring apoA I and apoB include chromatography, electrophoresis, and immunoassay. Among them, chromatography and electrophoresis are cumbersome to operate, and they cannot perform batch sample analysis and directly go to full-automatic biochemical analyzers. The immunodiffusion method, radioimmunoassay method, fluorescent labeling immunoassay method, enzyme-labeled immunoassay method, and chemiluminescence immunoassay method also have many deficiencies. If special equipment is required, the sample needs to be pretreated, and it cannot be used on a global scale. Automatic biochemical analyzer for batch detection and analysis, etc.

The immunoturbidimetric method commonly used in clinical practice is popular because of its small amount of sample, which can be directly analyzed in batches on an automatic biochemical analyzer, and is easy to operate. However, the currently established methods and reagents have some shortcomings and / or Insufficient, mainly manifested in: First, the antibody titer is not high, all below 1:16, and the specificity, affinity, and affinity are not good; second, the calibration serum is based on human serum, although the hepatitis B surface antigen is negative, HIV detection Negative human serum with normal alanine aminotransferase, but it is difficult to rule out the possibility of no other infectious diseases, which brings great danger to the operator, and it is a freeze-dried product, and the value of each batch is different , needs to be reconstituted with water before use, the operation is extremely inconvenient, and the reconstitution of different reconstituters varies greatly between the bottles. After reconstitution, it must be frozen and stored, especially if it cannot be thawed repeatedly, which will not only cause waste of reagents, but also affect the quality. To ensure that the measurement results vary greatly, and its stability is not good; the third is that the anti-interference ability of high-fat, jaundice, and hemolysis samples is poor

Fourth, the single-point calibration solution is used for calibration, which does not conform to the calculation of the curve regression equation, resulting in the phenomenon of low high values ​​and high low values, and the measured results are inaccurate

Fifth, the antiserum precipitates in a short period of time, making it difficult to operate on the machine and affecting the detection effect

The disadvantage of the above method is that the ultracentrifugation takes a long time, and the protein is often denatured and the spatial structure of the protein is changed during the process of degreasing and ultracentrifugation, and the immunogenicity of the protein is changed, so that the antiserum effect of the immune obtained The price is not high