Comparative analysis of fusion tags used to functionalize recombinant antibodies

In this work we evaluated the impact on expression yields, shelf-stability, thermostability and binding affinity of a set of C-terminal tags fused to the same anti-Her2 nanobody

Gianluca Veggiani

2019

Key concepts

Scholarcy highlights

  • Recombinant antibodies can be expressed as fusion constructs in combination with tags which simplify their engineering into reliable and homogeneous immunoreagents by allowing site-specific, 1:1 functionalization
  • In this work we evaluated the impact on expression yields, shelf-stability, thermostability and binding affinity of a set of C-terminal tags fused to the same anti-Her2 nanobody
  • We assessed the efficiency of the derivatization process
  • The constructs always bore a 6xHis tag plus either the controls or CLIP, HALO, AviTag, the LEPTG sequence recognized by Sortase A, or a free cysteine
  • The advantages and drawbacks of the different systems were analyzed and discussed

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