Assessing four subdomain-specific affinity resins' capability to separate half-antibody from intact bispecific antibody

Protein Expr Purif. 2022 Oct:198:106124. doi: 10.1016/j.pep.2022.106124. Epub 2022 May 31.


Half-antibody is a frequent byproduct associated with the recombinant production of many asymmetric bispecific antibodies (bsAbs). Although this byproduct can be largely removed by post-capture polishing steps, it is ideal to have it partially cleared at the capture step to achieve a more robust downstream process. Previously we showed that Protein A affinity chromatography possesses the capability to separate half-antibody. In this study, we assessed the half-antibody separation capability of four less commonly used subdomain-specific affinity resins. The data suggest that these resins exhibit different capabilities for separating half-antibody from the corresponding bsAb. In specific, whereas Protein A affinity resin can always provide partial separation under typical conditions, the separation efficiency of three subdomain-specific affinity resins (i.e., Capto L, CaptureSelect CH1-XL and CaptureSelect FcXP) heavily relies on the property of the two parental monospecific antibodies from which the bsAb is derived, which may range from complete separation to no separation at all. This novel information provides more options for half-antibody clearance at the capture step.

Keywords: Bispecific antibody (bsAb); Capto L; CaptureSelect CH1-XL; CaptureSelect FcXP; Half-antibody; KappaSelect; MabSelect SuRe LX; Protein A.

MeSH terms

  • Antibodies, Bispecific* / chemistry
  • Staphylococcal Protein A


  • Antibodies, Bispecific
  • Staphylococcal Protein A