Table 1 Nanobody sequence, specificity and neutralisation activity.

From: Nanobodies mapped to cross-reactive and divergent epitopes on A(H7N9) influenza hemagglutinin using yeast display

Nb

Sequence

Binding specificity (inactivated whole virus antigen)

Neutralisation

VHH CDR3 sequence (family)

TX/2012 (H3N2)

M/N/2000 (H7N3)

NY/2003 (H7N2)

AN/2013 (H7N9)

GD/2016 (H7N9)

HK/2017 (H7N9)

TX 2012 (H3N2)

SH 2013 (H7N9)

NB7-01

ALARSVDRLTYDY (1)

-

2.83

2.53

2.26

2.59

1.94

-

++++

NB7-02

HASPRDVLVDYDY (2)

-

1.98

2.44

1.63

2.48

0.26

-

++

NB7-03

AADFLTLQDMCVFSPITRYHD (3)

-

2.04

-

1.30

-

0.52

-

+++

NB7-04

AASPIPDLRDYDY (4)

-

0.84

1.62

0.71

3.37

0.31

-

 +

NB7-05

NVWIQFHNY

-

0.17

-

0.19

-

0.38

-

-

NB7-07

AFGATLYGVRRDEYAY

-

1.99

1.96

1.46

2.73

0.48

-

+++

NB7-08

AADRYSGPYDLGTISPRQDHYDV

-

0.91

1.52

0.91

2.65

2.05

-

-

NB7-09

ALARSVDRLEYDW (1)

-

2.86

2.82

2.37

3.30

2.60

-

++++ +

NB7-10

AADDLTLQDMCVMSPISRYHY (3)

-

1.64

-

1.20

-

2.30

-

+++

NB7-11

AVARSVDRLEYDY (1)

-

1.03

1.23

1.24

3.12

2.53

-

++++

NB7-12

HASPRDVLVDYD (2)

-

1.73

2.57

1.78

3.28

2.47

-

++++

NB7-13

AADFLTLQDMCVLSPITRYRS (3)

-

1.14

-

0.72

-

2.37

-

+++

NB7-14

ALARSVDRLTYDY (1)

-

2.67

2.75

2.08

3.00

2.57

-

+++++ 

NB7-15

AAGRTYCGLRGYEYDY

-

2.23

0.20

1.92

2.97

2.56

-

++++ 

NB7-16

AASPIPTLRDYDY (4)

-

1.44

1.71

1.08

2.86

2.42

-

+++

NB7-17

AASPRRLMCVPAEFDLYY

-

2.44

2.57

1.97

2.39

2.07

-

++++ 

NB37X-01

AQERERIGTTIRYY

0.36

0.13

0.22

0.48

0.38

2.14

-

-

NB37X-02

AAPKTRYLAPRAMESEFDY (5)

2.72

-

0.12

-

0.26

2.48

-

-

NB37X-03

AAPKTRYLAPRMTESDYDY (5)

0.12

-

-

-

0.34

-

-

-

NB37X-04

NALCYGCTPSSY

0.30

0.28

-

0.14

0.49

2.83

-

-

NB37X-05

AANPGTVVGRTRLSPRLRVPDEYDS

0.40

0.14

0.17

0.26

0.31

2.32

-

-

NB37X-06

AADRVDYPGGGATCRTSSAAYDH

0.26

-

0.20

0.20

0.36

2.55

-

-

  1. VHH-CDR3 sequences and family grouping of related sequences. ELISA is given as the average OD450nm reading of two wells coated with whole virus antigen and Nbs added at a single concentration of 5 µg/ml. – indicates OD450 < 0.1 and represents no binding. Inactivated whole virus antigens used TX/2012(H3N2) [A/Texas/50/2012 (H3N2)], M/N/2000(H7N3) [A/Mallard/Netherlands/12/2000], NY/2003 (H7N2) [A/New York/107/2003], AN/2013 (H7N9) [A/Anhui/01/2013], GD/2016 (H7N9) [A/Guangdong/17SF003/2016], HK/2017 (H7N9) [A/HongKong/125/2017]. Pseudotype based micro-neutralisation assays using A/Texas/50/2012 (H3) and A/Shanghai/02/2013 (H7) HA bearing pseudoviruses. += 500-3000 nM, ++ < 500 nM, +++< 100 nM, ++++  < 50 nM, +++++< 10 nM, - no neutralisation.
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