What are the methods of antibody preparation

There are various immunization routes, such as intravenously, intraperitoneally, intramuscularly, intradermally, subcutaneously, and intra-lymph node injections, etc. Subcutaneous or dorsal multi-point intradermal injections are commonly used, with each point injected with about 0.1ml. The choice of route is determined by the biological and physicochemical properties of the antigen, such as hormones, enzymes, toxins and other biologically active antigens, generally not suitable for intravenous injection. Rabbits are the animals most commonly used to produce antibodies, so the collection of rabbit blood is discussed here. For larger animals such as sheep, blood is collected from the jugular vein and artery, and for small animals such as rats, the relevant information can be consulted. There are two ways to collect rabbit blood, one is to bleed from the marginal ear vein or ear artery, and the other is to enter the carotid artery, or to collect blood from the heart. When taking arterial or venous bloodletting, the rabbit is put into a special wooden box or cage, with the ear exposed outside the box (cage), or the rabbit body can be caught by another person. Cut off the hair at the edge of the ear and apply a little xylene to the ear. After 30s, the ear blood vessels are dilated and congested. Pull the tip of the ear gently with the hand, and use a one-sided razor or a pointed surgical blade to cut the artery or vein quickly, and the blood will flow out, and 30~40 ml can be collected each time. then stop the bleeding by pressing with a cotton ball, and wash off the xylene after coagulation. After two weeks, blood can be bled in the other ear. This method can be repeated many times bloodletting. When bloodletting the carotid artery, put the rabbit on its back, fix it on the rabbit platform, cut off the hair of the neck, cut the skin, expose the carotid artery, insert the tube and bleed it. The process of bloodletting should be carried out in strict accordance with the requirements of asepsis.

Collected blood was placed at room temperature for about 1h, coagulated, placed at 4℃, overnight (do not freeze) precipitated serum, centrifuged, 4000rpm, 10min. in aseptic conditions, aspirated serum, divided (0.05~0.2ml), stored in the refrigerator at -40℃ or freeze-dried stored in the refrigerator at 4.C. for preservation.