Analytica chimica acta, 2008-08, Vol.623 (1), p.66-75
2008
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- Autor(en) / Beteiligte
- Titel
- Monitoring of influenza virus hemagglutinin in process samples using weak affinity ligands and surface plasmon resonance
- Ist Teil von
- Analytica chimica acta, 2008-08, Vol.623 (1), p.66-75
- Ort / Verlag
- Amsterdam: Elsevier B.V
- Erscheinungsjahr
- 2008
- Quelle
- MEDLINE
- Beschreibungen/Notizen
- Surface plasmon resonance (SPR) was used to screen the interaction between a variety of affinity ligands and hemagglutinin (HA) from human influenza virus, with the aim of identifying low affinity ligands useful for the development of a rapid bioanalytical sensor. Three sialic acid-based structures and four lectins were evaluated as sensor ligands. The sialic acid-based ligands included a natural sialic acid-containing glycoprotein, human α1-acid glycoprotein (α1-AGP), and two synthetic 6′-sialyllactose-conjugates, with varying degree of substitution. The interaction of HA with the four lectin-based ligands, concanavalin A (Con A), wheat germ agglutinin (WGA), Maackia amurensis lectin (MAL), and Sambucus nigra agglutinin (SNA), showed a wide variation of affinity strengths. Affinity and kinetics data were estimated. Strong affinities were observed for Con A, WGA, α1-AGP, and a 6′-sialyllactose-conjugate with a high substitution degree, and low affinities were observed for MAL and a 6′-sialyllactose-conjugate with low substitution. The main objective, to identify a low affinity ligand which could be used for on-line monitoring and product quantification, was met by a 6′-sialyllactose–ovalbumin conjugate that had 0.6 mol ligand per mol carrier protein. The apparent affinity of this ligand was estimated to be 1.5 ± 0.03 μM ( K D) on the SPR surface. Vaccine process samples containing HA were analyzed in the range 10–100 μg HA mL −1 and correlated with single-radial immunodiffusion. The coefficient of variation on the same chip was between 0.010 and 0.091.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 0003-2670, 1873-4324eISSN: 1873-4324DOI: 10.1016/j.aca.2008.06.005Titel-ID: cdi_swepub_primary_oai_DiVA_org_lnu_1816
- Format
- –
- Schlagworte
- Affinity ligand, Analytical chemistry, Animals, Binding Sites, Biotechnology, Bioteknik, Carrier Proteins - metabolism, Cells, Cultured, Chemistry, Exact sciences and technology, General, instrumentation, Hemagglutinin Glycoproteins, Influenza Virus - chemistry, Hemagglutinin Glycoproteins, Influenza Virus - metabolism, Humans, Influenza A Virus, H1N1 Subtype - immunology, Influenza A Virus, H1N1 Subtype - metabolism, Influenza A Virus, H3N2 Subtype - chemistry, Influenza A Virus, H3N2 Subtype - immunology, Influenza Vaccines - biosynthesis, Influenza Vaccines - metabolism, Influenza Vaccines - standards, Influenza virus, Influenza virus hemagglutinin, Kinetics, Lectins - metabolism, Ligands, Low affinity, Models, Molecular, N-Acetylneuraminic Acid - chemistry, N-Acetylneuraminic Acid - metabolism, NATURAL SCIENCES, NATURVETENSKAP, Orthomyxoviridae - immunology, Orthomyxoviridae - metabolism, Ovalbumin - chemistry, Ovalbumin - metabolism, Ovum - immunology, Ovum - metabolism, Protein Binding, Protein Conformation, Sambucus nigra, Substrate Specificity, Surface plasmon resonance, Surface Plasmon Resonance - methods, Time Factors, Weak affinity