Micro scale self-interaction chromatography of proteins: A mAb case-study

• Published in: Journal of Chromatography A Vol. 1434; pp. 57 - 63
• Main Authors: Hedberg, S.H.M., Heng, J.Y.Y., Williams, D.R., Liddell, J.M.
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 19.02.2016
• Subjects: Animals; Antibodies, Monoclonal - chemistry; Automation; Chickens; Chromatography; Chromatography, Liquid - instrumentation; Chromatography, Liquid - methods; Formulation; Liquid chromatography; Monoclonal antibodies; Muramidase - chemistry; Protein stability; Proteins; Scale-down; Screening; Second virial coefficient; Self-interaction chromatography; Monoclonal antibodies; Second virial coefficient; Scale-down; Protein stability; Self-interaction chromatography; Formulation
• ISSN: 0021-9673; 1873-3778
• DOI: 10.1016/j.chroma.2015.12.034

•Describes a methodology for micro scale SIC using 10 times less protein.•Examine a number columns of different sizes/aspect ratios.•Calculated B22 using a variety of conditions for all the columns for mAbs.•Similar results were obtained for all the different columns.•Guidance given on amount of proteins needed for different SIC scales. Self-interaction chromatography is known to be a fast, automated and promising experimental technique for determination of B22, but with the primary disadvantage of needing a significant amount of protein (>50mg). This requirement compromises its usage as a technique for the early screening of new biotherapeutic candidates. A new scaled down SIC method has been evaluated here using a number of micro LC columns of different diameters and lengths, using typically 10 times less stationary phase than traditional SIC. Scale-down was successfully accomplished using these micro-columns, where the SIC results for a range of differing columns sizes were in agreement, as reflected by k′, B22 and column volumes data. The results reported here demonstrate that a scaled down version of SIC can be easily implemented using conventional liquid chromatography system where the final amount of mAbs used was 10 times less than required by conventional SIC methodologies.