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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.