Small molecule clearance in ultrafiltration/diafiltration in relation to protein interactions: Study of citrate binding to a Fab

Ultrafiltration/diafiltration (UFDF) is commonly utilized in the purification of recombinant proteins to concentrate and buffer exchange the product. It is often the final step in the purification process, placing the protein in its final formulation and clearing small molecules introduced in upstre...

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Bibliographic Details
Published in:Biotechnology and bioengineering 2009-04, Vol.102 (6), p.1718-1722
Main Authors: Harinarayan, C, Skidmore, K, Kao, Y, Zydney, A.L, van Reis, R
Format: Article
Language:English
Subjects:
Acetates - chemistry
Algorithms
Antigens
binding
Binding sites
Biological and medical sciences
Biotechnology
citrate
Citric Acid - chemistry
clearance
diafiltration
formulation
Fundamental and applied biological sciences. Psychology
HEPES - chemistry
Humans
Immunoglobulin Fab Fragments - chemistry
Models, Chemical
Molecules
Monoclonal antibodies
NMR
Nuclear Magnetic Resonance, Biomolecular
Protein Interaction Domains and Motifs
Proteins
Recombinant Proteins - chemistry
small molecules
Tromethamine - chemistry
ultrafiltration
Ultrafiltration - methods
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Summary:Ultrafiltration/diafiltration (UFDF) is commonly utilized in the purification of recombinant proteins to concentrate and buffer exchange the product. It is often the final step in the purification process, placing the protein in its final formulation and clearing small molecules introduced in upstream purification steps. This article presents a case study of reduced small molecule clearance in ultrafiltration/diafiltration of an antigen-binding fragment of a monoclonal antibody. Citrate, a commonly utilized small molecule in downstream processes, is shown to have reduced clearance due to specific interactions with the protein product. The study presents process solutions and utilizes a simple model to characterize clearance of small molecules which exhibit interactions with product protein. Biotechnol. Bioeng. 2009;102: 1718-1722.
ISSN:0006-3592
1097-0290
DOI:10.1002/bit.22196