Monitoring of influenza virus hemagglutinin in process samples using weak affinity ligands and surface plasmon resonance
| Autor: | Carl-Fredrik Mandenius, Anna Aldén, Sabine Thébault, Gunnar Bergström, Charles Lutsch, Ronghui Wang, Sten Ohlson |
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| Rok vydání: | 2008 |
| Předmět: |
Models
Molecular Time Factors Ovalbumin Protein Conformation education Orthomyxoviridae Hemagglutinin (influenza) Hemagglutinin Glycoproteins Influenza Virus Ligands medicine.disease_cause Biochemistry Virus Substrate Specificity Analytical Chemistry chemistry.chemical_compound Influenza A Virus H1N1 Subtype Lectins Influenza A virus medicine Animals Humans Environmental Chemistry Binding site Surface plasmon resonance Cells Cultured Spectroscopy Ovum Binding Sites biology Chemistry Influenza A Virus H3N2 Subtype Lectin Surface Plasmon Resonance biology.organism_classification N-Acetylneuraminic Acid Sialic acid Kinetics Influenza Vaccines biology.protein Biophysics Carrier Proteins Protein Binding |
| Zdroj: | Analytica Chimica Acta. 623:66-75 |
| ISSN: | 0003-2670 |
| DOI: | 10.1016/j.aca.2008.06.005 |
| Popis: | 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 alpha1-acid glycoprotein (alpha1-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, alpha1-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 microM (K(D)) on the SPR surface. Vaccine process samples containing HA were analyzed in the range 10-100 microg HA mL(-1) and correlated with single-radial immunodiffusion. The coefficient of variation on the same chip was between 0.010 and 0.091. |
| Databáze: | OpenAIRE |
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