Application of mixed-mode resins in the purification of collagen

Aug 08,2025

Bestchrom

Bestchrom Mixed-mode resins combine ion exchange chromatography , hydrophobic interaction and hydrogen interaction chromatography interactions. In light of its excellent resolution and high tolerance towards high conductivity in sample loading due to unique mechanism, the type of reins are usually used in the effective isolation of recombinant collagens from clarified yeast or animal cell culture harvest, as well as the collagen isolation from E.coli lysate. This article will focus on the application of mixed-mode resins in the purification of recombinant collagens.

Resin selection strategy for collagen purification

Fig.1  Resin selection strategy for collagen purification

Introduction of Mixed-mode MMC resins

Bestchrom launched two MMC resins: Diamond MMC and Diamond MMC Mustang.

Table 1. Diamond MMC和Diamond MMC Mustang technical parameters 

After comparing the two resins, Diamond MMC has bigger particle size and higher ionic capacity, which can be used for the purification of collagen from clarified feedstock. Diamond MMC Mustang has smaller particle size and higher resolution, which can be used in the initial capture or polishing of collagens from collagen degraded fragments. The two resins can tolerate sample loading at high salinity condition.

Applications

• Case 1: Diamond MMC resin used for the purification of yeast-expressed collagen III

Sample：conductivity 35.16mS/cm，pH3.0

Fig.2 Diamond MMC chromatogram

In the case study, severe degradation was observed in target molecules. After optimizing conductivity and pH in washing and elution steps, removal of degraded segments and miscellaneous proteins was achieved, getting collagens with purity of 94.95%.

• Case 2: Diamond MMC Mustang used in the purification of yeast-expressed collagen XVII

Sample：Use water to dilute(dilution ration is 2)，adjust conductivity to 21.23mS/cm，pH4.0

Fig.3 Diamond MMC Mustang chromatogram

In the case, conductivity is 21.23mS/cm at sample loading with target protein flowing through. Two-step washing can achieve good results, collagen of high purity is finally achieved via linear elution.

Conclusion

Based on its multiple interactions, Mixed-mode chromatography resins enjoy merits such as high salinity tolerance in sample loading, and high resolution. The resins are used in one-step chromatography process of collagen, as well as the polishing of two-step chromatography process of collagen. As the recombinant collagen sector develops, mixed-mode resins are enjoying increasing significance in its process due to its high efficiency and flexibility.

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