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Table 1 Average relative rankings of scFv expression levels in various E. coli compartments and levels of display on filamentous phage.

From: Single-chain Fv phage display propensity exhibits strong positive correlation with overall expression levels

  Average scFv Rankings Major Germline Families
Clone Supernatant Periplasm Spheroplast Display VH VL
C1 6.0 ± 1 9.8 ± 0.8 7.0 ± 1.2 11.7 ± 1.1 VH4 VL3
C2 6.0 ± 0 14.3 ± 0.7 3.0 ± 0.6 6.2 ± 0.5 VH1 VL3
C3 7.5 ± 0.5 3.5 ± 0.5 8.7 ± 2.9 6.3 ± 1.0 VH4 VK1
C4 14.5 ± 0 11.8 ± 1.2 4.0 ± 1.5 4.2 ± 1.2 VH4 VK1
C5 14.5 ± 0 6.0 ± 1 3.7 ± 1.5 2.7 ± 0.3 VH4 VK1
N1 10.0 ± 2 13.8 ± 3.5 10.0 ± 0.6 7.5 ± 2.2 VH2 VK1
N2 4.0 ± 0 4.5 ± 2.2 3.3 ± 1.9 2.8 ± 1.1 VH1 VK1
N3 13.0 ± 0 10.8 ± 0.5 12.0 ± 0 11.5 ± 0.9 VH3 VK1
N4 9.0 ± 0 3.5 ± 0.5 9.3 ± 2.0 12.2 ± 1.1 VH3 VL2
N5 8.3 ± 3.3 4.3 ± 2.7 11.0 ± 1.0 10.7 ± 1.6 VH3 VL1
J1 10.5 ± 0.5 9.0 ± 3 14.7 ± 0.3 12.2 ± 1.1 VH3 VL2
J2 2.0 ± 0 11.8 ± 1.8 4.0 ± 1.5 3.8 ± 0.5 VH1 VK1
J3 2.0 ± 1 3.8 ± 2.2 5.3 ± 1.3 6.5 ± 1.3 VH3 VL2
J4 2.0 ± 1 3.5 ± 1.5 10.0 ± 1.0 11.3 ± 0.7 VH6 VL2
J5 10.8 ± 0.7 10.0 ± 1 14.0 ± 0.6 11.3 ± 1.5 VH3 VL2
  1. Densitometry data derived from the western blots shown in Figures 1 and 2 were used to rank scFvs from 1 to 15 in order of highest to lowest levels of scFv respectively. Rankings from repeat experiments were averaged and are shown here along with standard error for the overall expression and display levels. The average relative spheroplast scFv level rankings were used as an indicator of overall expression level and to designate the scFvs into three groups of either "high (average ranking ≤ 5; bold numbers)", "medium (average ranking ≥ 5 but ≤ 10; italicised numbers) " or "low" (average ranking ≥ 11; No Shading) expression. Rankings for phage display propensity were also used to assign similar groupings. The major VH and VL germline designations for each scFv are also shown for each clone.