Can pH of a His buffer change during lyophilization?

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SUMMARY

The discussion centers on the pH change observed in a 10mM histidine (His) buffer during the lyophilization of a 50mg/ml monoclonal antibody (mAb) solution. Initially, the buffer had a pH of 6.2, but upon reconstitution, the pH increased to 6.5. Participants noted that histidine buffers are generally less susceptible to pH changes during lyophilization compared to phosphate buffers. However, the specific composition of the His buffer, including the starting materials and pH adjustment methods, is crucial for understanding the observed pH shift.

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  • Understanding of histidine buffer preparation and its components.
  • Knowledge of lyophilization processes and their effects on protein stability.
  • Familiarity with pH measurement techniques and implications for protein formulations.
  • Basic concepts of monoclonal antibody (mAb) stability and formulation.
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Researchers, biochemists, and pharmaceutical scientists involved in protein formulation, particularly those working with monoclonal antibodies and lyophilization techniques.

Dhh1994
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We lyophilized 50mg/ml mAb with PI of 7.5 in a 10mM His buffer pH 6.2. Upon reconstitution the pH is 6.5. Does anyone have an explanation for this? I am under the impression His is much less susceptible to pH change during lyophilization as opposed to something like a phosphate buffer. Thanks.
 
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I cannot answer from direct experience or knowledge. A quick look showed that there is quite a lot of literature on stability and lyophilisation of proteins, naturally. I would think that this is a small pH change is not a worry in itself. For average readers of this forum, mAb probably means monoclonal antibody.

To attempt to reason about it (no promise) it is not enough to tell us ‘His’ (histidine) buffer. You would have to tell us how it is made up. E.g. whetherstarting with histidine or histidine hydrochloride or other salt, and brought to pH with what?
 
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