Elution mobile phase and process for immunoaffinity chromatography of viruses
Inventors
Brgles, Marija • Halassy, Beata • Forcic, Dubravko
Assignees
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Publication Number
US-10597640-B2
Publication Date
2020-03-24
Expiration Date
Abstract
The present invention discloses a novel composition of the elution mobile phase for virus purification by immunoaffinity chromatography which is consisting of one or more amino acids: L-serine, L-asparagine, or L-glutamine, or their salts with pharmaceutically acceptable acids; one or more auxiliary ingredients: L-arginine, glycine or imidazole, or their salts with pharmaceutically acceptable acids; one or more pharmaceutically acceptable pH adjusting agents for correcting the pH value of the mobile phase from pH=6.0-8.0; and purified water, up to 100% w/w of the mobile phase composition. The invention provides the use of immunoaffinity chromatography as a key step in the production of viral vaccines and/or viral vectors, in separation of infectious from non-infectious viral particles, and for enrichment of the viral suspension in infectious viral particles.
Core Innovation
The invention relates to an elution mobile phase for immunoaffinity chromatography of viruses and viral vectors. The elution mobile phase is a solution comprising one or more amino acids selected from L-serine, L-asparagine, and L-glutamine, one or more ingredients selected from L-arginine, glycine, imidazole, or their salts with pharmaceutically acceptable acids, one or more pharmaceutically acceptable pH adjusting agents, and purified water, with the pH corrected to 6.0–8.0.
The disclosed immunoaffinity purification process uses a covalently bound virus-specific antibody to form an immunoaffinity chromatography column on a chromatographic stationary phase. The column is equilibrated with a binding buffer, contaminated viruses are loaded, contaminants and non-bonded viruses are washed out, and purified virus bounded on the stationary phase is eluted using the claimed elution mobile phase to obtain an eluate as a suspension of purified virus in the mobile phase.
The document states that the method enables separation or enrichment of infectious versus non-infectious viral particles while preserving infectivity, and that native or physiological pH elution is used. Development studies using ovalbumin are described as showing improved native-pH elution yields with L-serine and combinations including L-serine with L-arginine and auxiliary ingredients such as imidazole, and experiments with mumps virus are described as showing higher infective recovery at native pH with the claimed amino-acid combinations compared with arginine-only elution at similar pH.
Claims Coverage
The partial document provides two independent claims that cover immunoaffinity chromatography-based virus isolation and purification and, separately, the elution mobile phase composition used for immunoaffinity elution. Across these claims, the mobile phase composition is defined by selected amino acids, specified auxiliary ingredients, pharmaceutically acceptable pH adjustment to pH 6.0–8.0, and purified water.
Immunoaffinity virus isolation and purification using covalently bound virus-specific antibody column with amino-acid elution mobile phase
A process for virus isolation and purification via immunoaffinity chromatography in which an antibody specific for a virus surface protein is covalently bound to a chromatographic stationary phase to form an immunoaffinity column, the column is equilibrated with a binding buffer and loaded with contaminated viruses, contaminants and non-bonded viruses are washed out, and the purified virus bounded on the stationary phase is eluted using an elution mobile phase comprising one or more amino acids selected from L-serine, L-asparagine, and L-glutamine, one or more ingredients selected from L-arginine, glycine, imidazole, pharmaceutically acceptable pH adjusting agents to correct pH to 6.0–8.0, and purified water.
Elution mobile phase for immunoaffinity chromatography of viruses defined by selected amino acids, auxiliary ingredients, and pH 6.0–8.0
A method for purification of viruses by using a mobile phase for elution in immunoaffinity chromatography of viruses, where the mobile phase is a solution comprising one or more amino acids selected from L-serine, L-asparagine, and L-glutamine, one or more ingredients selected from L-arginine, glycine, imidazole, one or more pharmaceutically acceptable pH adjusting agents to pH 6.0–8.0, and purified water.
Taken together, the independent claims cover immunoaffinity chromatography-based virus isolation and purification and the specific aqueous elution-mobile-phase composition used for immunoaffinity elution. Both claims hinge on selected amino acids, specified auxiliary ingredients, pharmaceutically acceptable pH adjustment to 6.0–8.0, and purified water; the process claim further specifies binding with a virus-specific covalently bound antibody column and elution yielding a suspension of purified virus.
Stated Advantages
Enables separation or enrichment of infectious versus non-infectious viral particles while preserving infectivity.
Improved native-pH elution yields are described in development studies.
Higher infective recovery at native pH is described for mumps virus with the claimed amino-acid combinations versus arginine-only elution at similar pH.
Documented Applications
Virus isolation and purification using immunoaffinity chromatography with a covalently bound virus-specific antibody column and the claimed elution mobile phase.
Purification of viral vectors by immunoaffinity chromatography using the mobile phase described.
Purification of viruses including Paramyxoviridae members such as mumps virus and measles virus, and other enumerated viruses such as Newcastle disease virus, human parainfluenza virus 1/2/3/4/5, Sendai virus, human metapneumovirus, and human respiratory syncytial virus.
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