We used AKTA prime machine to purify the serum and now we need to choose which one of the techniques above mentioned are most suitable for further use. To purify and measure the quantity of Ab in solution obtained from AKTA.
Any suggestions?
Immunodiffusion and agglutination techniques are modest in vitro techniques for detecting antibodies, but better and more sensitive techniques include Enzyme immunoassays, immunofluorescence assays and Chemiluminescence assays. If you want to purify antibodies from serum, the a viable approach could be by DEAE-52 column chromatography followed by affinity chromatography using for example, activated Sepharose 4-B and other related gels.There are plenty of protocols for these on the net.
Agglutination and immunodiffusion techniques mostly useful to check the antigen antibody specificty and interactions. The earlier one with insoluble antigens (cells) and later with soluble antigen (proteins in solution). It is difficult to employ any one of that to purify either antibodies or antigens. However, the best way to have one in pure form to prepare your own affinity matrix to purify the other from crude mixture. That is antibody coupled column for antigen purification from cell homogenates or specific antigen coupled for antibody purification from immune sera.
The agglutination and immunidiffusion can not be used for purification. It is widely used properly for detection of the antibody.
The agglutination and immune-diffusion can not be used for purification. It is widely used for detection of specific antibody/antigen in immune-diagnosis/prognosis of diseases or disorders particularly for to those infectious diseases where diagnosis by culture is difficult or not possible with in 24-48h.
The two methods you indicated are not meant for purification of antoibdies, since there are many other ways to puirify antibody or antibodies currently available.,
@shuen What if other methods are not available ? can these methods be use then ?
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