Physical and chemical analysis:

In the early stage of antibody drug discovery, physicochemical analysis is a crucial step to determine whether the candidate molecule can be eventually approved as a drug. In view of this, Sanyou Bio has established an comprehensive physicochemical analysis platform.

Instruments:

Sanyou Bio possess a lot of equipments for physicochemical analysis, including Agilent 1100 High Performance Liquid Chromatography (HPLC), ABI 7500 Fast RT-PCR, Bio-rad Protein Electrophoresis, Bacterial Endotoxin Thermostat, and so on.

Indicators Detection contents Analysis method
Stability Denaturation temperature measurement DSF
Purity and molecular weight Identification of Molecular Weight and Purity SDS-PAGE
Accurate identification of purity SEC
Safety Bacterial endotoxin test LAL
Titer Relative Identification of Concentration ProteinA-Titer
Charge heterogeneity Charge heterogeneity analysis CEX

Case study:

Aggregation analysis and detection by SEC (TSK3000 columne)

Sample Peak Area of polymer % Peak area of monomer % Peak area of low molecular weight substances %
Sample 1 2.2 76.0 21.8

CONCLUSION: The method can accurately separate high and low molecular weight substances, achieve the purpose of aggregation analysis and provide accurate data support for purity identification of proteins.

DSF(Tm detection)

CONCLUSION: This method can accurately detect the Tm value of the antibody drugs listed on the market, which is consistent with the Tm value reported in the literature, and the method is stable and reliable.

In the early stage of antibody drug screening, the early screen-ing of candidates antibody affinity kinetics is helpful for the project to rapidly narrow the range of candidate molecules, which is also a key step to rapidly promote the project of antibody drug development. In view of this, sanyou set up a comprehensive biochemical analysis platform, committed to the development of preferred antibodies.

Instruments:

Methods:

Affinity ranking


Blocking assay

Conclusion: Affinity ranking by ELISA at the Fab level provides a quick guide for subsequent molecule selection.

Conclusion: Candidate molecules with blocking activity equal to or better than the control antibody by ELISA could be selected. (The dotted red line represents the control antibody)

      Kientics detection

Conclusion: This method can accurately detect the affinity betweenthe antibody and the corresponding antigenupto2 . 23nM , and accurately show the Kon and Koff data, providing strong data support forthenext step of antibody optimization.

Technical process:

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