| Outline of Final Research Achievements |
We developed a digestion enzyme-immobilized monolith for the continuous fragmentation and separation of fragmented antibodies. Following the column, we connected two types of affinity columns, Protein A and Protein L. Excess fragments with a molecular weight below 25,000 bypass the purification column, while fragmented antibodies or unreacted antibodies could be recovered from either of the columns. Using polyclonal antibodies as a substrate, we investigated the effects of column residence time, pH, and temperature on fragmented antibody yield. The results indicated that while the reaction yield is dependent on the column residence time, high-temperature conditions can improve the yield even with shorter residence times.
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