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. 2023 Feb 20;15(1):2177978. doi: 10.1080/19420862.2023.2177978

Figure 3.

Fab-phage production with E. coli SK25. A shows immunoblots of SpyDisplay phages displaying Fabs of different specificities (adalimumab or trastuzumab), produced either with VCSM13 or Hyperphage as helper phage. Both specificities show comparable display rates, with a much higher level of display for the Fab-phages produced with Hyperphage. B shows the use of ELISA to test the binding of polyvalent Fab-phages to cognate and irrelevant antigens. The results confirm a highly target specific binding of the Fab-phages and no binding to the negative controls. C shows the elution of monovalent Fab-phages from MaxiSorp plates after treatment with TEV Protease. The results indicate an elution efficiency of around 90% for both specificities tested.

Production of Fab-phages.

(a) Immunoblots of SpyDisplay phages displaying Fabs of adalimumab (A) or trastuzumab (T) and produced with VCSM13 or Hyperphage as helper phage. Detection was performed with anti-M13-pIII followed by sheep anti-mouse IgG (H/L):HRP or anti-FLAG-HRP. Bands corresponding to the heavy chain fusion, degraded heavy chain fusion, and wildtype pIII are marked. (b) ELISA with polyvalent Fab-phages on cognate and irrelevant antigens, detection with anti-pVIII-HRP. (c) Fractions of monovalent Fab-phages eluted from MaxiSorp plates after treatment with TEV Protease for 30 minutes, relative to buffer control.