Oral avian interferon fusion protein and applications of oral avian interferon fusion protein as immunopotentiators
A technology of immune enhancer and fusion protein, which is applied in the field of oral poultry interferon fusion protein and its application as an immune enhancer, which can solve the problems of immune failure, enhanced vaccination response, increased side effects, etc., and is simple and easy to operate Effect
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0074] Embodiment 1, the preparation of fusion protein
[0075] 1. Construction of recombinant plasmids
[0076] 1. Synthesize the double-stranded DNA molecule shown in sequence 2 of the sequence listing.
[0077] The double-stranded DNA molecule shown in Sequence 2 of the sequence listing is named ChIFN-α-β fusion gene.
[0078] The double-stranded DNA molecule shown in sequence 2 of the sequence listing encodes the protein shown in sequence 1 of the sequence listing. The protein shown in Sequence 1 of the sequence listing is named ChIFN-α-β fusion protein.
[0079] In sequence 1 of the sequence listing, amino acid residues 1-193 constitute chicken interferon-alpha, amino acid residues 194-203 constitute a connecting peptide (flexible Linker), and amino acid residues 204-406 constitute chicken interferon-beta .
[0080] In Sequence 2 of the sequence listing, the 1st-579th nucleotides encode chicken interferon alpha, the 580th-609th nucleotides encode the connecting peptid...
Embodiment 2
[0128] Embodiment 2, the mensuration of antiviral activity
[0129] 1. Take well-grown DF-1 cells, suspend them in DMEM medium containing 10% FBS after digestion, and obtain a cell concentration of 5×10 5 cells / mL of cell suspension.
[0130] 2. Take a 96-well cell culture plate, add the cell suspension (100 μl / well) prepared in step 1, and incubate at 37°C, 5% CO 2 Cultivate under conditions for 8-10h (form monolayer cells).
[0131] 3. Take the his prepared in step 4 of Example 1 6 - ChIFN-α-β solution, which is diluted with DMEM culture solution containing 10% FBS to obtain a solution with a protein concentration of 0.001 mg / ml, which is named mother solution. The mother liquor was taken, and 4-fold gradient dilution was performed with DMEM culture fluid containing 10% FBS to obtain 6 gradient dilutions.
[0132] 4. Take the 96-well plate that completed step 2, suck and discard the culture supernatant, add 10% FBS-containing DMEM culture solution (100 μl / well) to 3 posi...
Embodiment 3
[0146] His in Example 3, Example 4, and Example 5 6 -The amount of ChIFN-α-β fusion protein is measured in U.
[0147] Embodiment 3, stability and shelf life are measured
[0148] Based on the particularity as an oral preparation, the His prepared in Step 4 of Example 1 6 -The stability of ChIFN-α-β solution (three production batches: LW2016001, LW2016002, LW2016003) under different conditions (especially the stability under high temperature and high humidity) was investigated.
[0149] Store under the condition of "temperature 4°C-8°C, relative humidity 60±10%" for 6 months, 12 months or 18 months, and then detect the protein titer according to the method of Example 2.
[0150] Store under the conditions of "temperature 25°C, relative humidity 60±10%" for 1 month, 3 months or 6 months, and then detect the protein titer according to the method in Example 2.
[0151] The results are shown in Table 2.
[0152] Table 2
[0153]
[0154] Stored under the conditions of "tem...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com