Avian glycan-specific IgM monoclonal antibodies for the detection and quantitation of type A and B haemagglutinins in egg-derived influenza vaccines

作者：Legastelois Isabelle^*; Chevalier Michel; Bernard Marie Clotilde; de Montfort Aymeric; Fouque Martine; Pilloud Alexandra; Serraille Christelle; Devard Nicolas; Engel Olivier; Sodoyer Regis; Moste Catherine

来源：Journal of Virological Methods, 2011, 178(1-2): 129-136.

DOI：10.1016/j.jviromet.2011.08.027

• Influenza
• IgM
• Monoclonal antibodies
• ELISA
• SRID
• Universal influenza reagent

Two IgM monoclonal antibodies (MAbs), Y6F5 and Y13F9, were selected during a screening of clones obtained immunising BALB/c mice with purified envelop proteins of the A/Sydney/5/97 (H3N2) IVR108 influenza strain. These MAbs recognised avian glycans on the haemagglutinin (HA) of the virus. This broad recognition allowed these MAbs to be used as enzyme-labelled secondary antibody reagents in a strain specific enzyme-linked immunosorbent assay (ELISA) in combination with a capture MAb that recognised and allowed the quantitation of the strain specific HA protein present in an egg-produced influenza vaccine. Advantage was taken of these MAbs to develop a universal ELISA in which the MAbs were used both as capture antibody and as enzyme-labelled secondary antibody to detect and quantify the HA protein of any egg-derived influenza vaccine. These avian-glycan specific IgM MAbs may prove to be particularly useful for determining the HA concentration in monovalent egg-derived pandemic influenza vaccines, in which the HA concentration may be lower than 5 mu g/ml. The HA detection limit in the ELISA assays developed in this study was 1.9 mu g/ml, as opposed to the 5 mu/ml quantitation limit generally accepted for the standard single-radial-immunodiffusion (SRID) assay, the approved technique for quantifying HA content in influenza vaccines. These ELISAs can also be used to quantify influenza HA formulated with emulsion-based or mineral salt adjuvants that could interfere with HA measurement by the SRID assay.

• 出版日期2011-12