title: Avian glycan-specific IgM monoclonal antibodies for the detection and quantitation of type A and B haemagglutinins in egg-derived influenza vaccines.
abstract: 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 5μg/ml. The HA detection limit in the ELISA assays developed in this study was 1.9μg/ml, as opposed to the 5μg/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.
authors: Department of Research, Sanofi Pasteur, 1541 Avenue Marcel Mérieux, 69280 Marcy L'Etoile, France. isabelle.legastelois@sanofipasteur.com
authors: Isabelle
authors: I
authors: Legastelois
authors: Michel
authors: M
authors: Chevalier
authors: Marie-Clotilde
authors: MC
authors: Bernard
authors: Aymeric
authors: A
authors: de Montfort
authors: Martine
authors: M
authors: Fouque
authors: Alexandra
authors: A
authors: Pilloud
authors: Christelle
authors: C
authors: Serraille
authors: Nicolas
authors: N
authors: Devard
authors: Olivier
authors: O
authors: Engel
authors: Régis
authors: R
authors: Sodoyer
authors: Catherine
authors: C
authors: Moste
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal; D000956:Antigens, Viral / Q000032:analysis* / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000032:analysis* / Q000276:immunology; D005779:Immunodiffusion / Q000379:methods; D007075:Immunoglobulin M; D009980:Influenza A virus / Q000254:growth & development* / Q000276:immunology; D007252:Influenza Vaccines / Q000737:chemistry* / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D011134:Polysaccharides / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D000956:Antigens, Viral; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D007075:Immunoglobulin M; D007252:Influenza Vaccines; D011134:Polysaccharides