Large-Scale Freezing of Biologics: Understanding Protein and Solute Concentration Changes in a Cryovessel—Part 2 - New data about cryoconcentration behavior at large scale. - BioPharm

ADVERTISEMENT

Large-Scale Freezing of Biologics: Understanding Protein and Solute Concentration Changes in a Cryovessel—Part 2
New data about cryoconcentration behavior at large scale.


BioPharm International
Volume 23, Issue 7

CONCLUSIONS

The results from mapping the frozen state suggest that cryoconcentration is preserved in the frozen state and the observed macro-cryoconcentration values during freezing were in general agreement with frozen state values. The extent of macro-cryoconcentration (measured by cores) varied between the top and bottom halves of the frozen block. The significantly higher macro-cryoconcentration in the lower half of the cryowedge can be explained by the presence of disaccharide in the formulation and formation of temperature-induced density gradients. When freezing a protein solution containing an additive that shows significant changes in density with temperature, a cooling-induced density gradient is created that leads to convective flow of solution. This causes the lower regions of the solution to cryoconcentrate to a greater extent than the upper regions. However, looking fundamentally at the changes occurring during the freezing process tells us that the true extent of (micro-)cryoconcentration experienced by the protein in the inter-dendritic spaces can be estimated from a phase or state diagram for the major solute component. This is significantly higher than what is measured as macro-cryoconcentration. Active freezing systems can affect the overall distribution of ice and solute in the matrix, i.e., the macro-cryoconcentration, but not the micro-cryoconcentration experienced by the solute and protein embedded in the matrix.

Parag Kolhe is a principal scientist, Alanta Lary is a senior scientist, Steven Chico is an associate scientist, Elisabeth Holding was an associate scientist, and Satish K. Singh is a research fellow, all at Pfizer, Inc., Chesterfield, MO, 636.247.9979,
.

REFERENCES

1. Tschoepe M, Schmidt R. Impact of freeze/thaw processing on monoclonal antibody stability. Bioprocess International Conference. Vienna, April 2008.

2. Kolhe P, Badkar A. Protein and solute distribution in drug substance containers during frozen storage as well as post-thawing: a tool to understand and fefine freezing-thawing parameters. Biotechnol Proc Devel. Submitted 2009.

3. Lashmar UT, Vanderburgh M, Little SJ. Bulk Freeze–Thawing of macromolecules: effects of cryoconcentration on their formulation and stability. Bioprocess Int. 2007;5(6):44–54.

4. Webb SD, Webb JN, Hughes TG, Sesin DF, Kincaid AC. Freezing biopharmaceutical using common techniques and the magnitude of bulk scale freeze-concentration. Bioprocess Int. 2002;15(5):22–34.

5. Kolhe P, Holding E, Lary A, Chico S, Singh SK. Large-scale freezing of biologics: understanding protein and solute concentration changes in a cryovessel–part I. BioPharm Int. 2010;23(6):53–60.

6. Zippelius R. Untersuchung zum Enifrier- und Aufauverhalten Pharmazeutischer Humanproteinlosungen im Grossmasstab. (Investigation of freezing and thawing process of human pharmaceutical protein solutions at large scale). PhD Thesis (in German), Ludwig-Maximilians University, Munich, 2002.

7. Wisniewski R, Wu VL. Large-Scale Freezing and Thawing of Biopharmaceutical Products. In: Avis KE, Wu VL, ed. Biotechnology and Biopharmaceutical Manufacturing, Processing, and Preservation Vol 2 of Drug manufacturing technology series: CRC Press, 1996:7–60.

8. Muldrew K, Acker JP, Elliott JAW, McGann LE. The water to ice transition: Implications for living cells. In: Fuller BJ, Benson EE, Lane N, editors. Life in the frozen state. Boca Raton, FL. CRC Press; 2004:67–108.

9. Roos Y, Karel M. Applying state diagrams to food processing and development. Food Technol. 1991 Dec;45(12):66, 68–71, 107.

10. Singh SK, Kolhe P, Wang W, Nema S. Large-scale freezing of biologics—a practitioner's review part 1: fundamental aspects. Bioprocess Int. 2009;7(10):32–44.


blog comments powered by Disqus

ADVERTISEMENT

ADVERTISEMENT

Pfizer to Acquire Vaccines from Baxter
July 30, 2014
GSK Submits EU Regulatory Filing for Malaria Vaccine Candidate
July 29, 2014
Bristol-Myers Squibb and Ono Pharmaceutical Collaborate on Immunotherapies
July 28, 2014
FDA Accepts First Biosimilar Filing
July 24, 2014
Compounding Pharmacy Issues Recall, But Challenges FDA Decision
July 22, 2014
Author Guidelines
Source: BioPharm International,
Click here