TSKgelChromatography column is used forADCDrug quality control analysis

Antibody conjugated drugs（ADC）It is a biopharmaceutical that couples anti-cancer drugs and other drugs into antibodies. In recent years, it has been used toADCThe research and development progress of new antibody drugs represented by bispecific antibodies is rapid, and currently there are16itemADCThe drug has been approved for marketing. In addition, R&D personnel have also begun to explore the third generation with higher uniformity and fewer side effects (off target toxicity)ADCMedication, thereforeADCThe purification and quality control analysis methods will become increasingly important.

1. ADCKey Quality Attributes and Their Analysis Methods
ADCThe key quality attributes and their analysis methods are shown in the table1As shown. The characteristic of non denaturing chromatography is that it can maintain the advanced structure and activity of antibodies during separation. Coupling ratio for separating drugs and antibodies(DAR)Hydrophobic chromatography is often used for different components.

In addition, volatile mobile phases can also be usedSEC/MSconductDARanalysis1）. As with antibodiesADCCThe separation associated with activity was also explored by the researchers through affinity chromatography for separationADC2，3）. There are also literature reports on antibodies and oligonucleotide conjugates(AOC)Anion exchange chromatography was used in the coupling ratio analysis 4）.

watch1. ADCKey Quality Attributes and Their Analysis Methods

TSKgelChromatography column is used forADCDrug quality control analysis

* MRM: Multiple reaction monitoring

Ref.: G. Petrie et al, White paper, 2017, https://www.eag 。。 ｃｏｍ/wp-content/uploads/2018/01/M-028517-Keys-to-Rapid-IND-Submission.pdf

2. ADCSeparation mode of non denaturing chromatographic analysis
analysisADCIn order to suppress the increased hydrophobicity caused by payload coupling, even under non denaturing conditions, it is sometimes necessary to add organic solvents such as isopropanol and acetonitrile to the mobile phase(5～25%).

watch2Summarized in the middleADCThe separation modes of non denaturing analysis and denaturing analysis, as well as their respective separation objects and characteristics.

watch2. ADCNon denaturing analysis/Separation mode of denaturing analysis

TSKgelChromatography column is used forADCDrug quality control analysis

3. ADCdrugSECanalysis
Size exclusion chromatography is commonly used for separationADCDimers, polymers, and fragments. The following image shows the use ofTSKgel UP-SW3000analysisADCThe spectrum.

picture2isADCDrugs are differentSECComparison of separation conditions on chromatographic columns. Compared to the other two modelsSECChromatographic column,TSKgel UP-SW3000More component information can be obtained in the separation of polymers.

4. ADCofDARValue analysis
When separating components with different coupling ratios of drugs and antibodies, high-resolution, non porous matrices are often usedHICChromatographic column. The new model recently listed by Dongcao CompanyHICchromatographic columnTSKgel HIC-ADC ButylimprovedADCThe separation selectivity can achieve more efficient analysis.

From the spectrum, it can be seen that,TSKgel HIC-ADC ButylseparationADCsimulant(DAR=0,2,4,6,8)The peak spacing is wider than other brands on the marketHICChromatographic columns have a wider separation range and better separation efficiency.