Preparation method of anti apoA1, apoB antibody and reagent box used for detecting apoAI, apoB

Abstract

A process for preparing the antibody of apoA I and apoB includes such steps as extracting the antigen apoA I and apoB, immunizing animal and purifying the antibody. A reagent kit for testing apoA I and apoB comprises antibody reagent, liquid serum type constant value calibrating liquid, and optional reactant. Their advantages are simple operation and process, high correctness, and high effect.

Description

technical field [0001] The invention relates to an antibody preparation method and a kit for measuring serum components, in particular to a preparation method for anti-APOAI and APOB antibodies and a kit for measuring apoAI and apoB in serum, which can be widely used in the technical fields of medicine and biochemistry. Background technique [0002] Apolipoprotein AI (apoAI) is the largest component of the apoA family. ApoAI 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 function of preventing atherosclerosis, therefore, measuring the content of apoAI 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 serum a...

AI Technical Summary

Problems solved by technology

[0003] Known methods for determining apoAI and apoB include chromatography, electrophoresis, and immunoassay. Among them, chromatography and electrophoresis are cumbersome to operate, and they cannot be analyzed in batches and directly on a fully automatic biochemical analyzer. The immunodiffusion method, radioimmunoassay method, fluorescent labeling immunoassay method, enzyme labeling 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 fully automated. 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, specificity, affinity, and affinity are not good, and 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