Prearation Of Affinity Filtration Carriers And Its Application In Purification Of Anti-hCG Antibody

At present, antibodies have been widely used in disease diagnosis,disease treatment and as carriers of targeted medicine. One of the mainreasons for its expensive price is the high cost of purifying. Although thereare many methods for purification of antibodies, there are still manyproblems such as the low resolution, low handling capacity and high cost ofthe medium. Therefore it is very necessary to develop a cheaper and moreconvenient method. In the paper, anti-HCG antibody and sulfamethazinewere used as a model protein and affinity ligand, respectively, the results ofstudy are as follows:First of all, dextran microspheres were used as carrier, sulfamethazinewas used as affinity ligand to prepare affinity filtration carrier, and thepreparation conditions were optimized. The ligand density was85.5μmol/g with high reuse ratio, ideal pH and mechanical stability. The purity andrecovery of affinity filtration were95%and69.8%, respectively.Sulfamethazine affinity carriers were used in affinity chromatograph,compared with protein A affinity chromatograph, the sulfamethazineaffinity chromatograph was better in high salt elution.Sulfamethazine and cellulose acetate membrane was used to prepareaffinity membrane, and the preparation conditions were optimized. Theresults showed that the ligand density is114.6μmol/g, the optimaladsorption pH was6.0, the maximum adsorption capacity was5.7mg/g, thepurity and recovery of affinity membrane separation was95%and37.6%,respectively.