Rapid purification of antiserum against Mycoplasma hyopneumoniae by an efficient absorption method.
A simple and efficient method for the removal of unwanted cross-reactive antibodies has been developed. The antiserum purification method was based on treatment of the antiserum with both sonicated extracts and boiling extracts of the Escherichia coli host cells used in immunoscreening the lambda EMBL3 library. We have demonstrated unambiguously that through this simple treatment, the rabbit anti-Mycoplasma hyopneumoniae antiserum can be effectively purified so that the amount of antibodies cross-reacted with Escherichia coli lysate proteins is drastically reduced. Compared with the traditional absorption methods, which require the chemical coupling of an absorbing agent to an insoluble support, and affinity purification methods, which have harsh denaturing condition, this method should greatly facilitate a successful immunoscreening experiment.
Division of Molecular Biology, Development Center For Biotechnology, Taipei, Taiwan, ROC.
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Chen
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Shiuan
D000818:Animals; D000907:Antibodies, Bacterial / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D007106:Immune Sera / Q000302:isolation & purification*; D007163:Immunosorbent Techniques; D009174:Mycoplasma / Q000276:immunology*; D011233:Precipitin Tests; D011817:Rabbits; D013997:Time Factors
D000907:Antibodies, Bacterial; D007106:Immune Sera