Skip to main content

Table 2 Potential or in-use quantification methods for influenza virus-like particles

From: Analytical technologies for influenza virus-like particle candidate vaccines: challenges and emerging approaches

 

Quantification method

Used with flu-VLPs

Sample type

LOD

LOQ

Working Range

SD (%RSD)

Assay Duration

Ref

HA antigen

SRID

Yes

P, NP,

~3 μg/ml

~3 μg/ml

3.12 – 100 ug/ml

(10%)

~30 h

[36]

HA assay

Yes

P, NP

2 HA unit/50ul

2 HA unit/50ul

2 - 2048 HA units/50 μl

~0,066 %

~5 h

[37, 38]

LC-MS

No

P

25 fmol

25 fmol

25 - 400fmol/10ul

(H1): 1.14 (3.5%)

n.s.

[39, 40]

(H3): 1.24 (5.6%)

(B): 5.04 (9.7%)

RP-HPLC

Yes

V

(H1): 0.25 ug/ml

(H1): 0.75ug/ml

2.5-100ug/ml

3.58 %

10-20 min

[41]

(B): 1 ug/ml

(B): 3ug/ml

7.73%

Universal ELISA

No

V

 

1ug/ml

0.2-16ug/ml

0.14-0.3 (14 - 30%)

n.s.

[42]

SPR immuno assays

No

P, NP, V

<0.5ug/ml

1ug/ml

1-15ug/ml

n.s.

n.s.

[43]

NA antigen

TBA NA assay

Yes

P

0.34 mU/ml

1.03 mU/ml

0.54 -6.47 mU/ml

2.95%

n.s.

[35]

NA star assay

Yes

P, NP

3 pM

3pM

3-10.5nM

n.s.

~1.5 h

[44]

Amplex Red NA assay

Yes

P

2.09 mU/ml

6.32mU/ml

2.00-7.00 mU/ml

16%

 

[35]

NA-Fluor Assay

Yes

P, NP

0.09 mU/ml

0.17-2.00 mU/ml

n.s.

2.55%

~4 h

[35]

HA & NA antigen

LC-MS

No

V

n.s.

n.s.

NA: 2-12ug/ml

n.s.

~5 h

[39]

HA: 10-70ug/ml

Universal SlotBlot

No

V, rHA, rNA

0.032 ug/ml

0.032 ug/ml

HA: 0.032-3ug/ml

2300 - 13500 (5%) particles/count

n.s.

[14, 45]

NA: 0.037-10 ug/ml

Total particles

DLS

No

P, NP

8.6 × 101 TCI50units/ml

8.6 × 101 TCI50units/ml

8.6 × 101 to 3.4 × 104 TCID50units/ml

n.s.

<30min

[46]

Virus Counter

No

P, NP

(NP): 3.6 × 106 VP/ml

(NP): 3.6 × 106 VP/ml

105 - 109 VP/ml

0.09 +/− 0.03 VP/ml

~40 min

[47]

(P): 1.0 × 105 VP/ml

(P): 1.0 × 105 VP/ml

AFFFF-MALS

No

P, NP

n.s.

n.s.

5 × 108 - 4.5 × 109 VP/ml

n.s.

~1 h

[48–50]

  1. Type of samples: P, Purified; NP, Non-purified; V, Vaccine doses; N.s, Non specified.