Development of an ELISA-Based Potency Assay for Inactivated Influenza Vaccines Using Cross-Reactive Nanobodies

被引：7

作者：

Cheung, Chung Y. ^{[1
,3
]}

Dubey, Sitara ^{[1
]}

Hadrovic, Martina ^{[1
,4
,5
]}

Ball, Christina R. ^{[2
]}

Ramage, Walter ^{[2
]}

McDonald, Jacqueline U. ^{[1
]}

Harvey, Ruth ^{[1
,6
]}

Hufton, Simon E. ^{[2
]}

Engelhardt, Othmar G. ^{[1
]}

机构：

[1] Med & Healthcare Prod Regulatory Agcy, Vaccines Div, Sci Res & Innovat, South Minams EN6 3QG, Potters Bar, England

[2] Med & Healthcare Prod Regulatory Agcy, Biotherapeut Div, Sci Res & Innovat, South Minams EN6 3QG, Potters Bar, England

[3] Univ British Columbia, Ctr Heart Lung Innovat, Vancouver, BC V6T 1Z4, Canada

[4] Univ Oxford, Dept Biol, Oxford OX1 3SZ, England

[5] Pirbright Inst, Ash Rd, Pirbright GU24 ONF, England

[6] Francis Crick Inst, Worldwide Influenza Ctr, London NW1 1AT, England

来源：

VACCINES | 2022年 / 10卷 / 09期

关键词：

influenza; vaccine; potency test; nanobodies; ELISA; cross-reactive; HEMAGGLUTININ; VIRUS; QUANTIFICATION; IMMUNOASSAY;

D O I：

10.3390/vaccines10091473

中图分类号：

R392 [医学免疫学]; Q939.91 [免疫学];

学科分类号：

100102 ;

摘要：

Inactivated vaccines are the main influenza vaccines used today; these are usually presented as split (detergent-disrupted) or subunit vaccines, while whole-virus-inactivated influenza vaccines are rare. The single radial immune diffusion (SRD) assay has been used as the gold standard potency assay for inactivated influenza vaccines for decades; however, more recently, various alternative potency assays have been proposed. A new potency test should be able to measure the amount of functional antigen in the vaccine, which in the case of influenza vaccines is the haemagglutinin (HA) protein. Potency tests should also be able to detect the loss of potency caused by changes to the structural and functional integrity of HA. To detect such changes, most alternative potency tests proposed to date use antibodies that react with native HA. Due to the frequent changes in influenza vaccine composition, antibodies may need to be updated in line with changes in vaccine viruses. We have developed two ELISA-based potency assays for group 1 influenza A viruses using cross-reactive nanobodies. The nanobodies detect influenza viruses of subtype H1N1 spanning more than three decades, as well as H5N1 viruses, in ELISA. We found that the new ELISA potency assays are sensitive to the nature of the reference antigen (standard) used to quantify vaccine antigens; using standards matched in their presentation to the vaccine type improved correspondence between the ELISA and SRD assays.

引用

页数：17

共 27 条

[1] Development of an enzyme-linked immunoassay for the quantitation of influenza haemagglutinin: an alternative method to single radial immunodiffusion [J].