Preparation method of apolipoprotein B antiserum

Preparation method of apolipoprotein B antiserum

  • CN 104,017,075 B
  • Filed: 01/06/2014
  • Issued: 02/28/2020
  • Est. Priority Date: 01/06/2014
  • Status: Active Grant
First Claim
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1. A method for preparing apolipoprotein B antiserum, which is characterized by comprising the following steps:

  • the method comprises the following steps;

    1) pretreatment of serum;

    treating serum for preparing apolipoprotein B by a polyanion compound with the concentration of 6-22% and a divalent metal ion with the concentration of 2-8mol/L to obtain low-density lipoprotein and very low-density lipoprotein;

    2) extraction of low density lipoprotein containing net apolipoprotein B;

    treating the obtained low-density lipoprotein by physiological saline and 0.5-1.1mol/L potassium oxalate in sequence, centrifuging, collecting supernatant, dialyzing to obtain crude low-density lipoprotein, density adjustment is carried out on the crude low-density lipoprotein by using a density adjusting reagent which is potassium bromide, performing ultracentrifugation by using a preparative ultracentrifuge after the adjustment of the density range of the low-density lipoprotein after being adjusted to be 1.03-1.05g/cm, the rotating speed of the ultracentrifugation is 40000-50000 r/min, the temperature is 8 ℃

    , then the low-density lipoprotein containing the apolipoprotein B is obtained through gel column chromatography, in the extraction process of the low-density lipoprotein net containing the apolipoprotein B, gel of gel column chromatography is agarose 4B gel column, and a substance of main peak chromatography is collected, namely the low-density lipoprotein net containing the apolipoprotein B antigen;

    3) preparation of apolipoprotein B antiserum;

    the apolipoprotein B is used as antigen to carry out immunoreaction to sheep to prepare apolipoprotein B antiserum, and the low-density lipoprotein which is acted on the sheep and contains the apolipoprotein B net has the density of 1.03-1.05g/cm and the dosage of 5-9mg per sheep.

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